鸭病毒性肿头出血症病毒感染特性及其免疫检测技术的研究
|
摘要
1998-2003年四川省、重庆市、贵州省和云南省等养鸭省份发生了一种以鸭头肿胀、眼结膜充血、出血,全身皮肤广泛性出血,肝肿胀呈土黄色,并伴有出血斑点,体温43℃以上,排草绿色稀粪等特征性的急性病毒性传染病。该病目前给养鸭业带来了巨大的经济损失。程安春等通过流行病学调查、临床症状和病理学变化观察、病理组织学检查、病原的分离鉴定、人工感染试验、电镜观察、防治试验,确诊为一种新的鸭病,并命名为“鸭病毒性肿头出血症(Duck viral swollen head hemorrhagic disease, DVSHD)”。国内其他学者也称之为“鸭传染性肿头症”或“鸭肿头败血症”。
本病与鸭瘟在临床特征和尸检变化上有许多相似之处,鸭病毒性肿头出血症病毒(Duck swollen head hemorrhagic disease virus, DSHDV)是该病的病原。目前有关该病的研究资料报道较少,特别是有关DSHDV感染特性以及特异、敏感、快速的检测方法等更是缺乏深入研究。因此,本文对鸭病毒性肿头出血症病毒的对成年鸭和鸭胚感染特性以及相关的免疫检测技术进行了一系列的研究。
本研究获得以下系列结果：
1.鸭病毒性肿头出血症实验感染模型的建立：本实验用鸭病毒性肿头出血症病毒强毒(HY-99株)作为种毒,通过肌注、口服和滴鼻三种方式感染28日龄健康鸭,攻毒后每天观察临床症状和发病鸭及死亡鸭的剖检变化。结果显示,感染鸭出现以体温升高、头颈肿胀、眼结膜充血、出血、排灰白色或草绿色稀粪为主要特征的临床症状和肝脏肿大、出血呈土黄色、免疫器官和消化道严重充血、出血为主要特征剖检变化。结果表明,临床症状和剖检变化与自然感染鸭基本一致,本实验成功构建了鸭病毒性肿头出血症实验感染模型。
2. DSHDV强毒人工感染鸭组织病理学发展规律研究：本研究对实验感染模型中感染鸭各个组织器官进行了动态组织病理学变化观察。结果表明,实验感染模型中三组鸭出现的组织病理学变化基本一致,但时间存在差异：肌注组鸭于感染后72h内、滴鼻组和口服组144h内组织病理学变化以充血、出血和炎性细胞浸润为主；肌注组鸭于感染72h后、滴鼻组和口服组144h后组织病理学变化以弥漫性出血、大小不等坏死灶或粘膜上皮细胞的坏死为主。免疫器官、消化腺、消化道、呼吸器官、心脏和肾脏等是主要的靶器官。这表明DSHDV是一种泛嗜性病毒,对全身各个组织器官具有严重破坏作用。
3. DSHDV强毒在人工感染鸭宿主细胞内的形态发生学和超微病理学研究：电镜下,完整DSHDV粒子呈圆形或椭圆形,无囊膜,直径为70nm-85nm。病毒粒子仅出现在感染细胞的胞浆中。电镜下,可观察到病毒粒子于感染4h后吸附于法氏囊靶细胞表面,并通过细胞的内吞作用进入细胞后,在胞浆内脱去双层衣壳,随后在胞浆内复制并合成病毒相关蛋白。这些合成结构蛋白、非结构蛋白及成熟或不成熟的病毒粒子在胞浆内聚集成圆形、椭圆形或不规则形的包涵体结构。感染后24h-72h,病毒核酸在这些基质上组装和成熟形成完整的病毒粒子。感染144h后,成熟病毒通过细胞的崩解而释放到细胞间隙。DSHDV强毒人工感染成年鸭后能够引起宿主细胞明显的超微结构变化(坏死和凋亡)。该病毒的主要靶器官包括法氏囊、胸腺、脾脏、肝脏、消化道和肾脏等器官。病毒侵染靶细胞主要有：淋巴细胞、肝细胞、成纤维细胞、巨噬细胞、上皮性网状细胞、粘膜上皮细胞、肠腺细胞等。
4. DSHDV强毒致人工感染鸭和鸭胚宿主细胞凋亡的研究：通过TEM法观察发现,DSHDV经尿囊腔感染10日龄鸭胚后尿囊膜上皮细胞、肝细胞、肠道上皮细胞和心肌细胞发生凋亡。凋亡细胞时间分布在感染后24h-96h,同时在部分宿主细胞的胞浆内发现少量病毒粒子。本研究利用HE染色法、TEM法和TUNEL法三种方法均观察到DSHDV感染诱导鸭宿主细胞的凋亡。发生凋亡的器官包括法氏囊、胸腺、脾脏、肾脏、肝脏、心脏、食管、腺胃和肠道。宿主细胞的凋亡指数从感染后2h到72h逐渐增加,72h达到最高值。感染72h后,濒死鸭和死亡鸭宿主细胞死亡以典型坏死为主。DSHDV感染诱导的细胞凋亡在DVSHD的致病机制中起到重要作用。
5.免疫酶组织化学检测DSHDV抗原方法的建立和应用：本研究以兔抗DSHDVIgG作为一抗、HRP-羊抗兔IgG作为二抗建立并优化了在石蜡切片中检测DSHDV抗原的间接免疫酶SP法,研究证实本方法具有较高的特异性和敏感性。该方法的应用研究揭示,三组鸭感染DSHDV后,肌注组(4h)、滴鼻组(12h)和口服组(24h)依次首先从法氏囊组织中检测阳性信号,随后抗原在多个组织中检出,包括免疫器官、消化腺、消化道、肾脏、心脏、肺脏和气管等。DSHDV侵染的靶细胞主要包括淋巴细胞、巨噬细胞、网状细胞、粘膜上皮细胞、肝细胞、间质细胞、腺泡细胞、肠腺细胞、肾小管上皮细胞、心肌细胞和柱状上皮细胞等。
6.间接ELISA检测鸭病毒性肿头出血症抗体方法的建立和应用：本研究以浓缩纯化的DSHDV作为包被抗原,建立了检测DSHDV抗体的间接酶联免疫吸附试验(ELISA)。试验最佳反应条件为：抗原最佳稀释度为1：40,高免血清为1：80稀释,最佳反应时间为60min；酶标抗体为1：2000稀释,最佳反应时间为60min；封闭液为1%BSA-PBST,封闭时间为60min；包被抗原最佳工作条件为37℃孵育1h后过夜；底物的最佳反应时间为37℃孵育15min。本实验建立的方法可成功应用于临床样品的检测和鸭免疫后抗体水平的评估。因此,本方法具有良好的特异性、稳定性、可重复性和较高的敏感性,可用于DVSHD抗体的检测。
An acute contagious viral disease in ducks that is characterized by swelling of the head; diarrhea with green dejecta; a hyperaemia and hemorrhagic of eyes conjunctiv; extensive hemorrhagic of skin; yellow and swollen liver with hemorrhagic; the high body temperature above 43℃, wes observed in many districts of Sichuan Province, Chongqing Municipality, Guizhou Province, and Yunnan Province, China, from 1998 to 2003. This disease has result in significant economic losses in the poultry industry as a result of high mortality and morbidity.This new duck disease was designated by Cheng et al as"duck viral swollen head hemorrhagic disease (DVSHD)", which based on epidemiological investigation, clinic signs, pathology changes, histopathological examination, isolation and identification of etiological agent, experimental infection, electron microscope observation and prevention and cure experiment. In addition, DVSHD was also named by "duck infectious swollen head" or "duck swollen head Septicemia".
Although there is some resemblance in clinic signs and autopsy between duck plague (DP) and DVSHD, remarkable differences are also observed in epidemic features and prevention and cure experiment between them.Duck swollen head hemorrhagic disease virus (DSHDV), which is considered as the causative agent of DVSHD, belongs to the Reoviridae family on the basis of its biological characteristis. The etiological agent of DVSHD was preliminarily determined, however, little information was available so far for the infection characteristics and the fast and effective methods for dectection of DSHDV or DVSHD. Therefore, a serial of experiments about them were carried out in this study.
The contents were summarized as follows:
1. Establishment of experimental infection model for DVSHD. In this study, 28-day-old normal ducks were infected with duck viral swollenhead haemorrhagic disease virus (DSHDV) virulent HY-99 strain by the way of intramuscular, oral and intranasal, respectively. After infection, the clinical symptoms and pathological changes of DSHDV-infected ducks were observed everyday. The results showed that, the DSHDV-infected ducks showed typical clinical symptoms, including fever, neck swelling, conjunctival hyperemia and hemorrhagic, and emitting white or grass green sparse muck; Necropsy revealed the typical lesions of DSHDV-infected ducks includes swelling, hemorrhagics, becoming soil-yellow of the liver, and severe hyperemia and hemorrhagic of the immune organs and digestive tract. The results suggested that the clinical symptoms and pathological changes of experimental DSHDV-infected ducks were consistent with natural DSHDV-infected ducks. So, the experimental model of DSHDV infection was successfully constructed in this experiment.
2. The histopathological studies on ducks experimentally infected with virulent DSHDV. In the basis of the above experimental infection model, study on the intimate histophological changes were performed in this research. The results indicated that ducks can fall to ill through the three routes and displayed similar histopathological changes but the infection time. Wthin 72h postinfection (PI) for intramuscular group,144h PI for intranasal group and oral group, the histopathological changes showed hyperemia and hemorrhagic and inflammatory cell infiltrating, and severe diffuse hemorrhagic, various degrees of degeneration, cytolysis and necrosis after 72h PI for intramuscular group,144h PI for intranasal group and oral group,. The critical organs contain immne organs, digestive gland, digestive tract, organum respiratorium, heart, and kidney and so on. These results indicated that DSHDV is a kind of pantropic virus and can result in severe lesion in every critical organ of duck.
3. Morphogenesis of virulent DSHDV and ultrastrural pathology of tissues in experimentally infected duck. The results of TEM observation showed that the complete DSHDV particles were round or ellipse, without peplos, with a diameter ranging from 75 nm to 85 nm, and they were presented in the cytoplasm of the infected cells. After the extracellular attachment of DSHDV to the host cell surface 12h PI, virions enter host cells by endocytosis and then DSHDV to uncoat in the cytoplasm.DSHDV replicate within the cytoplasm and synthesize viral proteins, which is formed spherical, ellipse or anomal inclsion., and then assemble within cytoplasmic inclusions from 24 to 72h PI. After 144h PI,.these complete and ripe virons were finally released to the extracellular space via the disruption of the cytoplasmic membrane. DSHDV infection can induce obviously ultrastructural changes, which mainly contain necosis and apoptosis. The target organs include bursa of Fabricius (BF), thymus, spleen, liver, digestive tract and kidney and so on. The target cells refer to lymphocytes, hepatocytes, desmocytes, macrophages, epithelial-reticular cells, mucosa endothelial cells and intestinal gland cells.
4. Study on host cell apoptosis induced by DSHDV in vivo. After 10-day-old duck e mbryo infected with virulent DSHDV by allantoic cavity, the results observed by TEM showed that DSHDV infection could induce host cell apoptsis,which contain chorioallantois endothelial cells, hepatocytes, intestinal tract endothelial cells and cadiocytes from 24 to 96h post infection(PI). Simultaneously, a few virus particles were observed within the cytoplasm. Virulent DSHDV infection could induce apoptosis in 28-day-old ducks through hematoxylin-eosin (HE) assay, transmission electron microscope (TEM) observation, and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labelling (TUNEL) assay. Apoptotic target organs refer to BF, thymus, spleen, kidney, liver, heart, oesophagus, proventriculus and intestinal tract. Apoptotic index (AI) values increased with time from 2 to 72 h PI, and the highest values were recorded at 72h PI. Further, cell death due to classic necrosis was observed in the dying or deceased ducks after 72h PI. In conclusion, host cell apoptosis can be induced by virulent DSHDV and may play an important role in the pathogenesis of DVSHD.
5. Development and application of an indirect immunoperoxidase assay for the detection of DSHDV antigen in tissues. An improved indirect immunoperoxidase assay (streptavidin-peroxidase assay, SP) was developed to virulent DSHDV antigens in paraformaldehyde-fixed, paraffin-embedded tissues of ducks, which based on rabbit antiDSHDV IgG as fisrst antibody and HRP labelled sheep anti rabbit IgG as second antibody. This assay in this study was proved to be highly specific and sensitive.The application of SP assay revealed that positive signals were first detected in BF at 4h PI for intramuscular injection group; 12h PI for nasal administration group and 24h PI for oral administration group, respectively. Next, DSHDV antigens were oberseved in more organs, which contains immune organs, digestive gland, digestive tract, kidney, heart, lung and trachea. The target cells refer to lymphocytes, macrophages, reticulocytes, mucosa epithelial-reticular cells, hepatocytes, mesenchymocyte, acinar cells, intestinal gland cells, renal tubular epithelial cells, cadiocytes and cellula columnoepithelialis.
6. Development and application of an indirect ELISA for the detection of antibodies against DSHDV. An indirect-ELISA for the detection and quantification of IgG antibody against DSHDV in serum was standardized using purified DSHDV as coating antigen. The test conditions were optimized by reagent titration utilizing known positive and negative sera. The best dilution was 1:40 for DSHDV antigen,1:80 for hyperimmune sera, and the best reaction tme for them 60min at 37℃. The best dilution and reaction time were respectively 1:2000 and 60min at 37℃. The best confining liquid and blockade time were respectively 1%BSA-PBST and 60min at 37℃. The best working condition for coating antigens was 4℃overnight after 37℃for 1h. Finally, the best reaction time for substrates was 37℃for 15min. The specificity test, sensitivity test and reproducibility test all proved that indirect ELISA had good specificity, stability, reproducibility and high sensitivity.This assay can be applied in the detection of antibody against DSHDV.
本病与鸭瘟在临床特征和尸检变化上有许多相似之处,鸭病毒性肿头出血症病毒(Duck swollen head hemorrhagic disease virus, DSHDV)是该病的病原。目前有关该病的研究资料报道较少,特别是有关DSHDV感染特性以及特异、敏感、快速的检测方法等更是缺乏深入研究。因此,本文对鸭病毒性肿头出血症病毒的对成年鸭和鸭胚感染特性以及相关的免疫检测技术进行了一系列的研究。
本研究获得以下系列结果：
1.鸭病毒性肿头出血症实验感染模型的建立：本实验用鸭病毒性肿头出血症病毒强毒(HY-99株)作为种毒,通过肌注、口服和滴鼻三种方式感染28日龄健康鸭,攻毒后每天观察临床症状和发病鸭及死亡鸭的剖检变化。结果显示,感染鸭出现以体温升高、头颈肿胀、眼结膜充血、出血、排灰白色或草绿色稀粪为主要特征的临床症状和肝脏肿大、出血呈土黄色、免疫器官和消化道严重充血、出血为主要特征剖检变化。结果表明,临床症状和剖检变化与自然感染鸭基本一致,本实验成功构建了鸭病毒性肿头出血症实验感染模型。
2. DSHDV强毒人工感染鸭组织病理学发展规律研究：本研究对实验感染模型中感染鸭各个组织器官进行了动态组织病理学变化观察。结果表明,实验感染模型中三组鸭出现的组织病理学变化基本一致,但时间存在差异：肌注组鸭于感染后72h内、滴鼻组和口服组144h内组织病理学变化以充血、出血和炎性细胞浸润为主；肌注组鸭于感染72h后、滴鼻组和口服组144h后组织病理学变化以弥漫性出血、大小不等坏死灶或粘膜上皮细胞的坏死为主。免疫器官、消化腺、消化道、呼吸器官、心脏和肾脏等是主要的靶器官。这表明DSHDV是一种泛嗜性病毒,对全身各个组织器官具有严重破坏作用。
3. DSHDV强毒在人工感染鸭宿主细胞内的形态发生学和超微病理学研究：电镜下,完整DSHDV粒子呈圆形或椭圆形,无囊膜,直径为70nm-85nm。病毒粒子仅出现在感染细胞的胞浆中。电镜下,可观察到病毒粒子于感染4h后吸附于法氏囊靶细胞表面,并通过细胞的内吞作用进入细胞后,在胞浆内脱去双层衣壳,随后在胞浆内复制并合成病毒相关蛋白。这些合成结构蛋白、非结构蛋白及成熟或不成熟的病毒粒子在胞浆内聚集成圆形、椭圆形或不规则形的包涵体结构。感染后24h-72h,病毒核酸在这些基质上组装和成熟形成完整的病毒粒子。感染144h后,成熟病毒通过细胞的崩解而释放到细胞间隙。DSHDV强毒人工感染成年鸭后能够引起宿主细胞明显的超微结构变化(坏死和凋亡)。该病毒的主要靶器官包括法氏囊、胸腺、脾脏、肝脏、消化道和肾脏等器官。病毒侵染靶细胞主要有：淋巴细胞、肝细胞、成纤维细胞、巨噬细胞、上皮性网状细胞、粘膜上皮细胞、肠腺细胞等。
4. DSHDV强毒致人工感染鸭和鸭胚宿主细胞凋亡的研究：通过TEM法观察发现,DSHDV经尿囊腔感染10日龄鸭胚后尿囊膜上皮细胞、肝细胞、肠道上皮细胞和心肌细胞发生凋亡。凋亡细胞时间分布在感染后24h-96h,同时在部分宿主细胞的胞浆内发现少量病毒粒子。本研究利用HE染色法、TEM法和TUNEL法三种方法均观察到DSHDV感染诱导鸭宿主细胞的凋亡。发生凋亡的器官包括法氏囊、胸腺、脾脏、肾脏、肝脏、心脏、食管、腺胃和肠道。宿主细胞的凋亡指数从感染后2h到72h逐渐增加,72h达到最高值。感染72h后,濒死鸭和死亡鸭宿主细胞死亡以典型坏死为主。DSHDV感染诱导的细胞凋亡在DVSHD的致病机制中起到重要作用。
5.免疫酶组织化学检测DSHDV抗原方法的建立和应用：本研究以兔抗DSHDVIgG作为一抗、HRP-羊抗兔IgG作为二抗建立并优化了在石蜡切片中检测DSHDV抗原的间接免疫酶SP法,研究证实本方法具有较高的特异性和敏感性。该方法的应用研究揭示,三组鸭感染DSHDV后,肌注组(4h)、滴鼻组(12h)和口服组(24h)依次首先从法氏囊组织中检测阳性信号,随后抗原在多个组织中检出,包括免疫器官、消化腺、消化道、肾脏、心脏、肺脏和气管等。DSHDV侵染的靶细胞主要包括淋巴细胞、巨噬细胞、网状细胞、粘膜上皮细胞、肝细胞、间质细胞、腺泡细胞、肠腺细胞、肾小管上皮细胞、心肌细胞和柱状上皮细胞等。
6.间接ELISA检测鸭病毒性肿头出血症抗体方法的建立和应用：本研究以浓缩纯化的DSHDV作为包被抗原,建立了检测DSHDV抗体的间接酶联免疫吸附试验(ELISA)。试验最佳反应条件为：抗原最佳稀释度为1：40,高免血清为1：80稀释,最佳反应时间为60min；酶标抗体为1：2000稀释,最佳反应时间为60min；封闭液为1%BSA-PBST,封闭时间为60min；包被抗原最佳工作条件为37℃孵育1h后过夜；底物的最佳反应时间为37℃孵育15min。本实验建立的方法可成功应用于临床样品的检测和鸭免疫后抗体水平的评估。因此,本方法具有良好的特异性、稳定性、可重复性和较高的敏感性,可用于DVSHD抗体的检测。
An acute contagious viral disease in ducks that is characterized by swelling of the head; diarrhea with green dejecta; a hyperaemia and hemorrhagic of eyes conjunctiv; extensive hemorrhagic of skin; yellow and swollen liver with hemorrhagic; the high body temperature above 43℃, wes observed in many districts of Sichuan Province, Chongqing Municipality, Guizhou Province, and Yunnan Province, China, from 1998 to 2003. This disease has result in significant economic losses in the poultry industry as a result of high mortality and morbidity.This new duck disease was designated by Cheng et al as"duck viral swollen head hemorrhagic disease (DVSHD)", which based on epidemiological investigation, clinic signs, pathology changes, histopathological examination, isolation and identification of etiological agent, experimental infection, electron microscope observation and prevention and cure experiment. In addition, DVSHD was also named by "duck infectious swollen head" or "duck swollen head Septicemia".
Although there is some resemblance in clinic signs and autopsy between duck plague (DP) and DVSHD, remarkable differences are also observed in epidemic features and prevention and cure experiment between them.Duck swollen head hemorrhagic disease virus (DSHDV), which is considered as the causative agent of DVSHD, belongs to the Reoviridae family on the basis of its biological characteristis. The etiological agent of DVSHD was preliminarily determined, however, little information was available so far for the infection characteristics and the fast and effective methods for dectection of DSHDV or DVSHD. Therefore, a serial of experiments about them were carried out in this study.
The contents were summarized as follows:
1. Establishment of experimental infection model for DVSHD. In this study, 28-day-old normal ducks were infected with duck viral swollenhead haemorrhagic disease virus (DSHDV) virulent HY-99 strain by the way of intramuscular, oral and intranasal, respectively. After infection, the clinical symptoms and pathological changes of DSHDV-infected ducks were observed everyday. The results showed that, the DSHDV-infected ducks showed typical clinical symptoms, including fever, neck swelling, conjunctival hyperemia and hemorrhagic, and emitting white or grass green sparse muck; Necropsy revealed the typical lesions of DSHDV-infected ducks includes swelling, hemorrhagics, becoming soil-yellow of the liver, and severe hyperemia and hemorrhagic of the immune organs and digestive tract. The results suggested that the clinical symptoms and pathological changes of experimental DSHDV-infected ducks were consistent with natural DSHDV-infected ducks. So, the experimental model of DSHDV infection was successfully constructed in this experiment.
2. The histopathological studies on ducks experimentally infected with virulent DSHDV. In the basis of the above experimental infection model, study on the intimate histophological changes were performed in this research. The results indicated that ducks can fall to ill through the three routes and displayed similar histopathological changes but the infection time. Wthin 72h postinfection (PI) for intramuscular group,144h PI for intranasal group and oral group, the histopathological changes showed hyperemia and hemorrhagic and inflammatory cell infiltrating, and severe diffuse hemorrhagic, various degrees of degeneration, cytolysis and necrosis after 72h PI for intramuscular group,144h PI for intranasal group and oral group,. The critical organs contain immne organs, digestive gland, digestive tract, organum respiratorium, heart, and kidney and so on. These results indicated that DSHDV is a kind of pantropic virus and can result in severe lesion in every critical organ of duck.
3. Morphogenesis of virulent DSHDV and ultrastrural pathology of tissues in experimentally infected duck. The results of TEM observation showed that the complete DSHDV particles were round or ellipse, without peplos, with a diameter ranging from 75 nm to 85 nm, and they were presented in the cytoplasm of the infected cells. After the extracellular attachment of DSHDV to the host cell surface 12h PI, virions enter host cells by endocytosis and then DSHDV to uncoat in the cytoplasm.DSHDV replicate within the cytoplasm and synthesize viral proteins, which is formed spherical, ellipse or anomal inclsion., and then assemble within cytoplasmic inclusions from 24 to 72h PI. After 144h PI,.these complete and ripe virons were finally released to the extracellular space via the disruption of the cytoplasmic membrane. DSHDV infection can induce obviously ultrastructural changes, which mainly contain necosis and apoptosis. The target organs include bursa of Fabricius (BF), thymus, spleen, liver, digestive tract and kidney and so on. The target cells refer to lymphocytes, hepatocytes, desmocytes, macrophages, epithelial-reticular cells, mucosa endothelial cells and intestinal gland cells.
4. Study on host cell apoptosis induced by DSHDV in vivo. After 10-day-old duck e mbryo infected with virulent DSHDV by allantoic cavity, the results observed by TEM showed that DSHDV infection could induce host cell apoptsis,which contain chorioallantois endothelial cells, hepatocytes, intestinal tract endothelial cells and cadiocytes from 24 to 96h post infection(PI). Simultaneously, a few virus particles were observed within the cytoplasm. Virulent DSHDV infection could induce apoptosis in 28-day-old ducks through hematoxylin-eosin (HE) assay, transmission electron microscope (TEM) observation, and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labelling (TUNEL) assay. Apoptotic target organs refer to BF, thymus, spleen, kidney, liver, heart, oesophagus, proventriculus and intestinal tract. Apoptotic index (AI) values increased with time from 2 to 72 h PI, and the highest values were recorded at 72h PI. Further, cell death due to classic necrosis was observed in the dying or deceased ducks after 72h PI. In conclusion, host cell apoptosis can be induced by virulent DSHDV and may play an important role in the pathogenesis of DVSHD.
5. Development and application of an indirect immunoperoxidase assay for the detection of DSHDV antigen in tissues. An improved indirect immunoperoxidase assay (streptavidin-peroxidase assay, SP) was developed to virulent DSHDV antigens in paraformaldehyde-fixed, paraffin-embedded tissues of ducks, which based on rabbit antiDSHDV IgG as fisrst antibody and HRP labelled sheep anti rabbit IgG as second antibody. This assay in this study was proved to be highly specific and sensitive.The application of SP assay revealed that positive signals were first detected in BF at 4h PI for intramuscular injection group; 12h PI for nasal administration group and 24h PI for oral administration group, respectively. Next, DSHDV antigens were oberseved in more organs, which contains immune organs, digestive gland, digestive tract, kidney, heart, lung and trachea. The target cells refer to lymphocytes, macrophages, reticulocytes, mucosa epithelial-reticular cells, hepatocytes, mesenchymocyte, acinar cells, intestinal gland cells, renal tubular epithelial cells, cadiocytes and cellula columnoepithelialis.
6. Development and application of an indirect ELISA for the detection of antibodies against DSHDV. An indirect-ELISA for the detection and quantification of IgG antibody against DSHDV in serum was standardized using purified DSHDV as coating antigen. The test conditions were optimized by reagent titration utilizing known positive and negative sera. The best dilution was 1:40 for DSHDV antigen,1:80 for hyperimmune sera, and the best reaction tme for them 60min at 37℃. The best dilution and reaction time were respectively 1:2000 and 60min at 37℃. The best confining liquid and blockade time were respectively 1%BSA-PBST and 60min at 37℃. The best working condition for coating antigens was 4℃overnight after 37℃for 1h. Finally, the best reaction time for substrates was 37℃for 15min. The specificity test, sensitivity test and reproducibility test all proved that indirect ELISA had good specificity, stability, reproducibility and high sensitivity.This assay can be applied in the detection of antibody against DSHDV.
引文
[1]Saif, Y.M.,Barnes, H.J.,Glisson, J.R. Disease of poultry[M].12th. Ames:Iowa State University Press,2008.
[2]苏敬良,黄瑜.新型鸭肝炎病毒的分离及初步鉴定[J].中国兽医科学,2002,32(1)：15-16.
[3]何冉娅,罗玉均,张桂红,等.一株新型鸭肝炎病毒的分离与初步鉴定[J].养禽与禽病防治,2008,(11)：14-16.
[4]范书才,李虹,袁率珍,等.新型鸭肝炎病毒的分离鉴定[J].中国预防兽医学报,2009,(10):770-775.
[5]Tseng, C.H.,Tsai, H.J. Molecular characterization of a new serotype of duck hepatitis virus[J]. Virus Res,2007,126 (1-2):19-31.
[6]Kim, M.C.,Kwon, Y.K.,Joh, S.J.et al. Recent korean isolates of duck hepatitis virus reveal the presence of a new geno-and serotype when compared to duck hepatitis virus type 1 type strains[J]. Arch Virol,2007,152 (11):2059-2072.
[7]刁有祥,吕桂霞,郑福英,等.一种新型鸭瘟病原的分离鉴定及其特性[J].中国兽医学报,2006,26(2)：136-139.
[8]Vickers, M.L.,Hanson, R.P. Newcastle disease virus in waterfowl in wisconsin[J]. J Wildl Dis,1982,18 (2):149-158.
[9]Spalatin, J.,Hanson, R.P.Epizootiogy of newcastle disease in waterfow[J]. Avian Dis,1975,19 (3):573-582.
[10]张训海,赵家龙.鸭副粘病毒强毒株的分离和鉴定[J].中国动物检疫,2001,18(10)：24-26.
[11]Liu, H.,Wang, Z.,Wang, Y.et al. Characterization of newcastle disease virus isolated from waterfowl in china[J]. Avian Dis,2008,52 (1):150-155.
[12]张太翔,田国宁,张金玲,等.鸭新城疫病毒强毒株的分离和初步鉴定[J].中国兽医杂志,2009,45(9)：35-37.
[13]Sandhu, T.S.,Calnek, B.W.,Zeman, L. Pathologic and serologic characterization of a variant of duck hepatitis type i virus[J]. Avian Dis,1992,36 (4):932-936.
[14]郑献进,张大丙,曲丰发,等.Ⅰ型鸭肝炎病毒变异株的鉴定[J].中国兽医杂志,2006,42(5)：15-16.
[15]柴同杰.山东省樱桃谷鸭群鸭圆环病毒及其混合感染的调查[J].中国家禽,2009,31(6)：4-7.
[16]Soike, D.,Albrecht, K.,Hattermann, K.et al. Novel circovirus in mulard ducks with developmental and feathering disorders[J]. Vet Rec,2004,154 (25):792-793.
[17]吴宝成,陈家祥,姚金水,等.番鸭呼肠孤病毒的分离与鉴定[J].福建农林大学学报(自然科学版),2001,30(2)：227-230.
[18]陈少莺,陈仕龙,林锋强,等.一种新的鸭病(暂名鸭出血性坏死性肝炎)病原学研究初报[J].中国农学通报,2009,25(16)：28-31.
[19]程安春,汪铭书,陈孝跃,等.一种新发现的鸭病毒性肿头出血症的研究[J].中国兽医科学,2003,33(10)：15-21.
[20]张舍郁,程安春,汪铭书,等.间接免疫荧光检测石蜡切片中鸭肿头出血症病毒及抗原定位方法的初步建立[J].中国农业科学,2008,41(3)：868-874.
[21]岳华,罗薇,汤承,等.鸭“传染性肿头症”的研究[J].四川畜牧兽医,2002,29(8)：24-25.
[22]盛敏.鸭肿头败血症病原特性研究[D].雅安：四川农业大学,2003.
[23]Kaleta, E.F. Herpesviruses of birds--areview[J]. Avian Pathol,1990,19 (2):193-211.
[24]Fauquet, C.M.,Mayo, M.A.,Maniloff, J. Virus taxonomy.Ⅷth report of the international committee on taxonomy of virus[M]. San Diego:Elsevier Academic Press,2005.
[25]Nii, S.,Uno, F.,Yoshida, M.et al. [structure and assembly of human beta herpesviruses][J]. Nippon Rinsho,1998,56 (1):22-28.
[26]Kocan, R.M. Duck plague virus replication in muscovy duck fibroblast cells[J]. Avian Dis,1976,20 (3):574-580.
[27]Wolf, K.,Burke, C.N.,Quimby, M.C. Duck viral enteritis:A comparison of replication by ccl-141 and primary cultures of duck embryo fibroblasts[J]. Avian Dis,1976,20 (3):447-454.
[28]Mettenleiter, T.C. Herpesvirus assembly and egress[J]. J Virol,2002,76 (4):1537-1547.
[29]Mettenleiter, T.C.,Klupp, B.G.,Granzow, H. Herpesvirus assembly:A tale of two membranes[J]. Curr Opin Microbiol,2006,9 (4):423-429.
[30]袁桂萍,程安春,汪铭书,等.鸭病毒性肠炎病毒强毒株的形态发生学与超微病理学研究[J].病毒学报,2004,20(4)：326-332.
[31]郭宇飞,程安春,汪铭书,等.鸭病毒性肠炎病毒CH强毒株在鸭胚成纤维细胞中形态发生学的研究[J]．畜牧兽医学报,2006,37(3)：274-280.
[32]Montali, R.J.,Bush, M.,Greenwell, G.A. An epornitic of duck viral enteritis in a zoological park[J].J Am Vet Med Assoc,1976,169 (9):954-958.
[33]Barr, B.C.,Jessup, D.A.,Docherty, D.E.et al. Epithelial intracytoplasmic herpes viral inclusions associated with an outbreak of duck virus enteritis[J]. Avian Dis,1992,36 (1):164-168.
[34]Davison, S.,Converse, K.A.,Hamir, A.N.et al. Duck viral enteritis in domestic muscovy ducks in pennsylvania[J]. Avian Dis,1993,37 (4):1142-1146.
[35]Campagnolo, E.R.,Banerjee, M.,Panigrahy, B.et al. An outbreak of duck viral enteritis (duck plague) in domestic muscovy ducks (cairina moschata domesticus) in illinois[J]. Avian Dis,2001,45 (2):522-528.
[36]彭广能,郭万柱,程安春,等.人工感染鸭瘟的病理学研究[J].中国兽医学报,2009,29(4)：445-450.
[37]Asplin, F.D. Duck hepatitis:Vaccination against two serological types[J]. Vet Rec,1965,77 (50):1529-1530.
[38]Haider, S.A.,Calnek, B.W. In vitro isolation, propagation, and characterization of duck hepatitis virus type iii[J]. Avian Dis,1979,23 (3):715-729.
[39]Tauraso, N.M.,Coghill, G.E.,Klutch, M.J. Properties of the attenuated vaccine strain of duck hepatitis virus[J].Avian Dis,1969,13 (2):321-329.
[40]Kim, M.C.,Kwon, Y.K.,Joh, S J.et al. Molecular analysis of duck hepatitis virus type 1 reveals a novel lineage close to the genus parechovirus in the family picornaviridae[J]. J Gen Virol,2006,87 (Pt 11):3307-3316.
[41]Tseng, C.H.,Knowles, N.J.,Tsai, H.J. Molecular analysis of duck hepatitis virus type 1 indicates that it should be assigned to a new genus[J]. Virus Res,2007,123 (2):190-203.
[42]Ding, C.,Zhang, D. Molecular analysis of duck hepatitis virus type 1[J]. Virology,2007,361 (1):9-17.
[43]Todd, D.,Smyth, V.J.,Ball, N.W.et al. Identification of chicken enterovirus-like viruses, duck hepatitis virus type 2 and duck hepatitis virus type 3 as astroviruses[J]. Avian Pathol,2009,38 (1):21-30.
[44]Toth, T.E. Chicken-embryo-adapted duck hepatitis virus growth curve in embryonated chicken eggs[J]. Avian Dis,1969,13 (3):535-539.
[45]郑献进.鸭肝炎病毒血清型鉴定及Ⅰ型变异株弱毒疫苗的研究[D].北京：中国农业大 学,2006.
[46]Hwang, J. Active immunization against duck hepatitis virus[J]. Am J Vet Res,1972,33 (12):2539-2544.
[47]Davis, D.,Woolcock, P.R. Passage of duck hepatitis virus in cell cultures derived from avian embryos of different species[J]. Res Vet Sci,1986,41 (1):133-134.
[48]Maiboroda, A.D. [cultivation of duck hepatitis virus in cell culture][J]. Veterinariia,1972,8: 50-52.
[49]Doroshko, I.N.,Bezrukavaia, I.,Golubnichii, V.P.et al. [production tests of a vaccine against viral hepatitis in ducks][J]. Veterinariia,1975, (1):52-54.
[50]Richter, W.R.,Rdzok, E.J.,Moize, S.M. Electron microscopy of viruslike particles associated with duck viral hepatitis[J]. Virology,1964,24:114-116.
[51]谢毅,邝荣禄,林维庆.鸭肝炎病毒形态及接种鸡胚引起的病理变化观察[J].华南农业大学学报,1989,10(4)：65-71.
[52]徐福南,周芳.鸭病毒性肝炎的组织病理学研究[J].中国兽医科学,1990,(2)：6-7.
[53]胡薛英,程国富.试验感染鸭病毒性肝炎雏鸭的组织病理学研究[J].华南农业大学学报,2000,19(1)：48-50.
[54]彭南秀.雏鸭病毒性肝炎的动态病理学研究[J].中国兽医科学,1998,28(8)：22-23.
[55]Farmer, H.,Chalmers, W.S.,Woolcock, P.R. The duck fatty kidney syndrome-an aspect of duck viral hepatitis[J]. Avian Pathol,1987,16 (2):227-236.
[56]陈建红,梁发朝,卢玉葵.鸭肝炎自然病例与鸭肝炎病毒人工发病病例组织病理学观察[J].中国预防兽医学报,2001,23(3)：203-205.
[57]祁保民,姚金水,卢惠明.鸭病毒性肝炎的病理形态学观察[J].福建农林大学学报(自然科学版),1999,28(3)：350-352.
[58]胡薛英,谷长勤,王德海,等.雏鸭胰脏组织损伤在新型鸭肝炎病毒感染过程中的变化特点[J].中国兽医学报,2006,26(2)：186-188.
[59]王永坤.禽流感流行新特点及防制对策[J].中国家禽,2002,24(12)：6-8.
[60]Fouchier, R.A.,Munster, V.,Wallensten, A.et al. Characterization of a novel influenza a virus hemagglutinin subtype (h16) obtained from black-headed gulls[J]. J Virol,2005,79 (5):2814-2822.
[61]Booy, F.P.,Ruigrok, R.W.,van Bruggen, E.F. Electron microscopy of influenza virus. A comparison of negatively stained and ice-embedded particles[J]. J Mol Biol,1985,184 (4)：667-676.
[62]田国斌,赵增连,唐秀英,等.鸭源禽流感病毒的分离和鉴定[J].中国预防兽医学报,1999,21(1)：1-3.
[63]李银,刘宇卓,张敬峰,等.一株鸭源H9亚型禽流感病毒在鸡胚上传代及繁殖规律的研究[J].江苏农业科学,2006,(4)：94-96.
[64]Briedis, D.J.,Conti, G.,Munn, E.A.et al. Migration of influenza virus-specific polypeptides from cytoplasm to nucleus of infected cells[J]. Virology,1981,111(1):154-164.
[65]Choppin, P.W.,Scheid, A. The role of viral glycoproteins in adsorption, penetration, and pathogenicity of viruses[J]. Rev Infect Dis,1980,2 (1):40-61.
[66]Krug, R.M.,Soeiro, R. Studies on the intranuclear localization of influenza virus-specific proteins[J].Virology,1975,64 (2):378-387.
[67]Compans, R.W.,Meier-Ewert, H.,Palese, P. Assembly of lipid-containing viruses[J]. J Supramol Struct,1974,2 (2-4):496-511.
[68]Franklin, R.M.,Hinnen, R.,Schafer, R.et al. Structure and assembly of lipid-containing viruses, with special reference to bacteriophage pm2 as one type of model system[J]. Philos Trans R Soc Lond B Biol Sci,1976,276 (943):63-80.
[69]李银.禽流感病毒H9N2鸭分离株的生物学与分子生物学特性及其快速诊断的研究[D].南京：南京农业大学,2006.
[70]刘超男,刘明,张云,等.H5N1亚型禽流感病毒的鸭致病性研究[J].中国农业科学,2006,39(2)：412-417.
[71]李玉谷,周泉鹤,崔聪颖,等.一株H5N1亚型高致病力禽流感病毒人工感染鸭的组织病理学观察[J].畜牧兽医学报,2008,39(10)：1373-1381.
[72]马春全,卢玉葵,邓桦,等.鸭H5N1型高致病性禽流感的病理组织学观察[J].中国兽医科学,2004,34(11)：11-14.
[73]岳华,刘群,汤承,等.“鸭传染性肿头症”病毒的分离及部分特性研究[J].西南民族学院学报,2002,28(3)：314-316.
[74]岳华,刘群,汤承,等.”鸭传染性肿头症”病毒对雏鸭致病性和卵黄抗体研究[J].西南民族学院学报,2002,28(4)：519-520.
[75]何平有,蒋文灿.实验性鸭肿头病败血症病理研究[J].黑龙江畜牧兽医,2006,11)：87-88.
[76]张舍郁,朱德康,程安春,等.鸭肿头出血症病毒强毒的致病性及感染组织细胞的超微病 理变化[J].中国兽医科学,2007,37(2)：1017-1023.
[77]张舍郁,程安春,汪铭书.鸭病毒性肿头出血症病毒培养特性与形态观察[J].黑龙江畜牧兽医,2007,(11)：9-11.
[78]杨汉春.动物免疫学[M].第二版.北京：中国农业大学出版社,2003.
[79]郭积燕.免疫学检验中的酶免疫技术[J].中华检验医学杂志,2005,28(2)：221-224.
[80]Voller, A.,Bidwell, D.E.,Bartlett, A. Enzyme immunoassays in diagnostic medicine. Theory and practice[J]. Bull World Health Organ,1976,53 (1):55-65.
[81]Nakane, P.K.,Pierce, G.B., Jr. Enzyme-labeled antibodies:Preparation and application for the localization of antigens[J]. J Histochem Cytochem,1966,14 (12):929-931.
[82]Avrameas, S.,Uriel, J. Method for labeling antigens and antibodies with enzymes and their application in immunodiffusion[J]. Compt Rend SerD,1966,262:2543-2545.
[83]Ram, J., S.,Nakane, P.K.,Rawlinson, D.G.et al. Enzyme-labelled antibodies for ultrastructural studies[J]. Fed Proc,1966,25:732.
[84]Miles, L.E.,Hales, C.N. Labelled antibodies and immunological assay systems[J]. Nature,1968,219 (5150):186-189.
[85]Avrameas, S. Coupling of enzymes to proteins with glutaraldehyde. Use of the conjugates for the detection of antigens and antibodies[J]. Immunochemistry,1969,6 (1):43-52.
[86]Engvall, E.,Perlmann, P. Enzyme-linked immunosorbent assay (ELISA). Quantitative assay of immunoglobuling[J]. Immunochemistry,1971,8 (9):871-874.
[87]Van Weemen, B.K.,Schuurs, A.H. Immunoassay using antigen-enzyme conjugates[J]. FEBS Lett,1971,15:232-235.
[88]Rubenstein, K.E.,Schneider, R.S.,Ullman, E.F. "Homogeneous" enzyme immunoassay. A new immunochemical technique[J]. Biochem Biophys Res Commun,1972,47 (4):846-851.
[89]Matsuoka, K.,Maeda, M.,Tsuji, A. Fluorescence enzyme immunoassay for insulin using peroxidase-tyramine-hydrogen peroxide[J]. Chem Pharm Bull (Tokyo),1979,27 (10):2345-2350.
[90]Wang, H.X.,Stott, R.A.,Thorpe, GH.et al. An enhanced chemiluminescence enzyme immunoassay for serum progesterone[J]. Steroids,1984,44 (4):317-328.
[91]Halsall, H.B.,Heineman, W.R. Electrochemical immunoassay:An ultrasensitive method[J]. J Int Fed Clin Chem,1990,2 (4):179-187.
[92]Alard, P.,Lantz, O.,Sebagh, M.et al. A versatile elisa-pcr assay for mma quantitation from a few cells[J].Biotechniques,1993,15 (4):730-737.
[93]Wu, M.,Stockley, P.G.,Martin, W.J.,2nd. An improved western blotting technique effectively reduces background[J]. Electrophoresis,2002,23 (15):2373-2376.
[94]Ahangaran, A.,Mohammadi, G.M.,Habibi, M.K.et al. Rapid diagnosis of soybean mosaic virus n soybean by tissue-print immunoassay and diba in comparison to other serological methods[J]. Commun Agric Appl Biol Sci,2006,71 (3PtB):1207-1212.
[95]钟晓琴,刘美英,黄云艳,等.一种基于酶标纳米金探针的蛋白质检测方法[J].中国医药生物技术,2009,4(3)：200-206.
[96]Sternberger, L.A. Immunocytochemistry[M].3rd. New York:John Wiley&Sons,1986.
[97]Maekawa, M. [properties of enzymes for enzyme immunoassay][J]. Nippon Rinsho,1995,53 (9):2154-2159.
[98]Krieg, R.,Halbhuber, K.J. Recent advances in catalytic peroxidase histochemistry[J]. Cell Mol Biol (Noisy-le-grand),2003,49 (4):547-563.
[99]Porstmann, T.,Kiessig, S.T. Enzyme immunoassay techniques. An overview[J]. J Immunol Methods,1992,150 (1-2):5-21.
[100]Ormanns, W.,Schaffer, R. An alkaline-phosphatase staining method in avidin-biotin immunohistochemistry[J]. Histochemistry,1985,82 (5):421-424.
[101]尹伯元,王仁芝,李振甲.标记免疫学[M].北京：原子能出版社,1998.
[102]Hancock, M.B. Leu-enkephalin, substance p, and somatostatin immunohistochemistry combined with the retrograde transport of horseradish peroxidase in sympathetic preganglionic neurons[J].J Auton Nerv Syst,1982,6 (2):263-272.
[103]Yamashita, S.,Korach, K.S. A modified immunohistochemical procedure for the detection of estrogen receptor in mouse tissues[J]. Histochemistry,1989,90 (5):325-330.
[104]Erdogan, D.,Elmas, C.,Ilgaz, C. Visualization of sperm by immunohistochemistry: Application of alternative counterstains[J]. Arch Androl,2005,51 (3):233-238.
[105]Chi, V.,Chandy, K.G. Immunohistochemistry:Paraffin sections using the vectastain abc kit from vector labs[J].J Vis Exp,2007, (8):308.
[106]Jiao, K.,Sun, W.,Zhang, S.S. Sensitive detection of a plant virus by electrochemical enzyme-linked immunoassay [J]. Fresenius J Anal Chem,2000,367 (7):667-671.
[107]Zelnik, V.,Harlin, O.,Fehler, F.et al. An enzyme-linked immunosorbent assay (ELISA) for detection of marek's disease virus-specific antibodies and its application in an experimental vaccine trial[J]. J Vet Med B Infect Dis Vet Public Health,2004,51 (2):61-67.
[108]Benbrook, D.M. An elisa method for detection of human antibodies to an immunotoxin[J]. J Pharmacol Toxicol Methods,2002,47 (3):169-175.
[109]Reyna-Bello, A.,Eleizalde, M.C.,Silva, A.M. Assessment of chromogen suitability in elisa for the detection of anaplasmosis and trypanosomosis[J]. J Immunoassay Immunochem,2007,28 (1):1-11.
[110]Khatun, S.,Nara, S.,Tripathi, Vet al. Development of elisa for measurement of progesterone employing 17-alpha-oh-p-hrp as enzyme label[J]. J Immunoassay Immunochem,2009,30 (2):186-196.
[111]Jiao, K.,YAO, H.,Xu, J.et al. Electrochemical studies of 1,5-dihydroxynaphthalene as substrate for voltammetric enzyme immunoassay[J]. Sci China Ser B,2004,47 (3):184-191.
[112]Jiao, G.Z.,Zhu, L.H.,Yao, H.et al. Voltammetric enzyme-linked immunoassay for dectection of horseradish peroxidase using phenol red as substrate [J]. J Qingdao Univ Sci & Tech,2004,25 (2):98-101.
[113]Zhang, S.S.,Jiao, K.,Chen, H.Y. Investigation of voltammetric enzyme-linked immunoassay based on a new system of oap-h2o2-hrp[J]. Electroanalysis,1999,11(7):511-516.
[114]Bobrow, M.N.,Harris, T.D.,Shaughnessy, KJ.et al. Catalyzed reporter deposition, a novel method of signal amplification. Application to immunoassays[J]. J Immunol Methods,1989,125 (1-2):279-285.
[115]Adams, J.C. Biotin amplification of biotin and horseradish peroxidase signals in histochemical stains[J]. J Histochem Cytochem,1992,40 (10):1457-1463.
[116]Mayer, G.,Bendayan, M. Biotinyl-tyramide:A novel approach for electron microscopic immunocytochemistry[J]. J Histochem Cytochem,1997,45 (11):1449-1454.
[117]Kohler, A.,Lauritzen, B.,Van Noorden, C.J. Signal amplification in immunohistochemistry at the light microscopic level using biotinylated tyramide and nanogold-silver staining[J]. J Histochem Cytochem,2000,48 (7):933-941.
[118]Hikino, T. In situ hybridization with novel biotinyl-tyramide:Fundamental studies and its utility of the detection of human papilloma virus in tissue sections[J]. Rinsho Byori,2007,55 (10):922-929.
[119]Krieg, R.,Halbhuber, K.J. Novel oxidative self-anchoring fluorescent substrates for the histochemical localization of endogenous and immunobound peroxidase activity[J]. J Mol Histol,2004,35 (5):471-487.
[120]沈关心,周汝霖.现代免疫学实验技术[M].武汉：湖北科学技术出版社,1998.
[121]Tang, B.,Wang, Y.,Du, M.et al. Catalytic spectrophotometric determination of ascorbic acid in tea drink with 1,5-bis(p-hydroxybenzaldene)thiocarbohydrazone as the substrate for horseradish peroxidase[J].JAgric Food Chem,2003,51 (15):4198-4201.
[122]Ichibangase, T.,Ohba, Y.,Kishikawa, N.et al. Chemiluminescence assay of lipase activity using a synthetic substrate as proenhancer for luminol chemiluminescence reaction[J]. Luminescence,2004,19 (5):259-264.
[123]Hainfeld, J.F. Enzymatic deposition and alteration of metals[P]. U.S. Patent,6670113,2003.
[124]Morgenstern, S.,Kessler, G.,Auerbach, J.et al. An automated p-nitrophenylphosphate serum alkaline phosphatase procedure for the autoanalyzer[J]. Clin Chem,1965,11 (9):876-888.
[125]Neumann, H.,van Vreedendaal, M. An improved alkaline phosphatase determination with p-nitrophenyl phosphate[J].Clin Chim Acta,1967,17 (2):183-187.
[126]Geneix, N.,Dufour, E.,Venien, A.et al. Development of a monoclonal antibody-based immunoassay for specific quantification of bovine milk alkaline phosphatase[J]. J Dairy Res,2007,74 (3):290-295.
[127]Anyaoku, V.N.,Wood, D.F.,Pavlou, R.et al. An enzyme-linked immunoadsorbent assay for human pituitary glycoprotein free alpha-subunit:Evaluation and initial experience in patients with pituitary disease[J].J Endocrinol,1993,136 (3):511-516.
[128]Avrameas, S.,Ternynck, T. Peroxidase labelled antibody and fab conjugates with enhanced intracellular penetration[J]. Immunochemistry,1971,8 (12):1175-1179.
[129]Sternberger, L.A. Immunocytochemistry[M].2nd. New York:John Wiley & Sons,1979.
[130]Imagawa, M.,Yoshitake, S.,Ishikawa, E.et al. Development of a highly sensitive sandwich enzyme immunoassay for human ferritin using affinity-purified anti-ferritin labelled with beta-d-galactosidase from escherichia coli[J]. Clin Chim Acta,1982,121 (3):277-289.
[131]Herrmann, J.E.,Morse, S.A. Coupling of peroxidase to poliovirus antibody:Characteristics of the conjugates and their use in virus detection[J]. Infect Immun,1973,8 (4):645-649.
[132]Wisdom, G.B. Conjugation of antibodies to horseradish peroxidase [J]. Methods Mol Biol,2005,295:127-130.
[133]Petrali, J.P.,Hinton, D.M.,Moriarty, GC.et al. The unlabeled antibody enzyme method of immunocytochemistry. Quantitative comparison of sensitivities with and without peroxidase-antiperoxidase complex[J]. J Histochem Cytochem,1974,22 (8):782-801.
[134]Sinclair, R.A.,Burns, J.,Dunnill, M.S. Immunoperoxidase staining of formalin-fixed, paraffin-embedded, human renal biopsies with a comparison of the peroxidase-antiperoxidase (pap) and indirect methods[J]. J Clin Pathol,1981,34 (8):859-865.
[135]Vacca, L.L.,Rosario, S.L.,Zimmerman, E.A.et al. Application of immunoperoxidase techniques to localize horseradish peroxidase-tracer in the central nervous system[J]. J Histochem Cytochem,1975,23 (3):208-215.
[136]Mason, D.Y.,Abdulaziz, Z.,Falini, B.et al. Single and double immunoenzymatic techniques for labeling tissue sections with monoclonal antibodies [J]. Ann N Y Acad Sci,1983,420:127-133.
[137]Moir, D.J.,Ghosh, A.K.,Abdulaziz, Z.et al. Immunoenzymatic staining of haematological samples with monoclonal antibodies [J]. Br J Haematol,1983,55 (3):395-410.
[138]Chilosi, M.,Lestani, M.,Pedron, S.et al. A rapid immunostaining method for frozen sections[J]. Biotech Histochem,1994,69 (4):235-239.
[139]Sabattini, E.,Bisgaard, K.,Ascani, S.et al. The envision++system:A new immunohistochemical method for diagnostics and research. Critical comparison with the apaap, chemmate, csa, labc, and sabc techniques[J]. J Clin Pathol,1998,51 (7):506-511.
[140]Shi, S.R.,Guo, J.,Cote, R.J.et al. Sensitivity and detection efficiency of a novel two-step detection system (powervision) for immunohistochemistry[J]. Appl. Immunohistochem. Mol. Morphol.,1999,7:201-208.
[141]Ogawa, T.,Gamoh, K.,Aoki, H.et al. Validation and standardization of virus neutralizing test using indirect immunoperoxidase technique for the quantification of antibodies to rabies virus[J]. Zoonoses Public Health,2008,55 (6):323-327.
[142]Hoffman, D.R. Enzyme-linked immunosorbent assays (ELISA) for immunoglobulin e and blocking antibodies[J]. Methods Enzymol,1981,73 (PtB):656-666.
[143]Hawkes, R.,Niday, E.,Gordon, J. A dot-immunobinding assay for monoclonal and other antibodies[J].Anal Biochem,1982,119 (1):142-147.
[144]Herbrink, P.,Van Bussel, F.J.,Warnaar,S.O. The antigen spot test (AST):A highly sensitive assay for the detection of antibodies [J]. J Immunol Methods,1982,48 (3):293-298.
[145]Karawajew, L.,Behrsing, O.,Kaiser, Get al. Production and elisa application of bispecific monoclonal antibodies against fluorescein isothiocyanate (FITC) and horseradish peroxidase (HRP)[J].J Immunol Methods,1988,111(1):95-99.
[146]Schwab, K.J.,McDevitt, J.J. Development of a PCR-enzyme immunoassay oligoprobe detection method for toxoplasma gondii oocysts, incorporating pcr controls [J]. Appl Environ Microbiol,2003,69 (10):5819-5825.
[147]Whiley, D.M.,Arden, K.E.,Mackay, I.M.et al. Simultaneous detection and differentiation of human polyomaviruses jc and bk by a rapid and sensitive PCR-elaha assay and a survey of the jcv subtypes within an australian population[J]. J Med Virol,2004,72 (3):467-472.
[148]Nguyen, T.D.,de Kesel, M.,Bigaignon, Get al. Detection of toxoplasma gondii tachyzoites and bradyzoites in blood, urine, and brains of infected mice[J]. Clin Diagn Lab Immunol,1996,3 (6):635-639.
[149]Zhou, M.,Liu, Q.,Wongsrichanalai, C.et al. High prevalence of plasmodium malariae and plasmodium ovale in malaria patients along the thai-myanmar border, as revealed by acridine orange staining and pcr-based diagnoses[J]. Trop Med Int Health,1998,3 (4):304-312.
[150]Bora, U.,Kannan, K.,Nahar, P. Heat-mediated enzyme-linked immunosorbent assay procedure on a photoactivated surface[J]. J Immunol Methods,2004,293 (1-2):43-50.
[151]Sharma, P.,Nahar, P. Ultrasound wave-mediated enzyme-linked immunosorbent assay technique[J].Anal Chim Acta,2009,650 (2):241-246.
[152]马吉飞,邱震东,郑兆荣,等.应用免疫组化PAP法检测兔出血症病毒抗原的动态分布[J].中国兽医学报,1994,14(3)：264-267.
[153]Halbur, P.G,Miller, L.D.,Paul, P.S.et al. Immunohistochemical identification of porcine reproductive and respiratory syndrome virus (PRRSV) antigen in the heart and lymphoid system of three-week-old colostrum-deprived pigs[J]. Vet Pathol,1995,32 (2):200-204.
[154]Last, R.D.,Jardine, J.E.,Smit, M.M.et al. Application of immunoperoxidase techniques to formalin-fixed brain tissue for the diagnosis of rabies in southern africa[J]. Onderstepoort J Vet Res,1994,61 (2):183-187.
[155]Clift, S.J.,Williams, M.C.,Gerdes, T.et al. Standardization and validation of an immunoperoxidase assay for the detection of african horse sickness virus in formalin-fixed, paraffin-embedded tissues[J]. J Vet Diagn Invest,2009,21(5):655-667.
[156]Carlsson, G.L.,Heller, R.S.,Serup, P.et al. Immunohistochemistry of pancreatic development in cattle and pig[J].Anat Histol Embryol,2010,39 (2):107-119.
[157]张晓梅,杨汉春,任慧英,等.用LSAB免疫组化染色法检测猪生殖-呼吸道综合征病毒抗原的研究[J].中国预防兽医学报,1999,21(6)：427-429.
[158]张平英,朱德康,程安春,等.间接免疫酶组织化学法检测猪繁殖与呼吸综合征病毒感染和抗原定位[J].中国兽医学报,2008,28(2)：128-132.
[159]Haines, D.M.,Moline, K.M.,Sargent, R.A.et al. Immunohistochemical study of hemophilus somnus, mycoplasma bovis, mannheimia hemolytica, and bovine viral diarrhea virus in death losses due to myocarditis in feedlot cattle[J]. Can Vet J,2004,45 (3):231-234.
[160]Njaa, B.L.,Clark, E.G,Janzen, E.et al. Diagnosis of persistent bovine viral diarrhea virus infection by immunohistochemical staining of formalin-fixed skin biopsy specimens[J]. J Vet Diagn Invest,2000,12 (5):393-399.
[161]Dougherty, R.M.,Marucci, A.A.,Distefano, H.S. Application of immunohistochemistry to study of avian leukosis virus[J].J Gen Virol,1972,15 (2):149-162.
[162]Islam, M.R.,Nessa, J.,Halder, K.M. Detection of duck plague virus antigen in tissues by immunoperoxidase staining[J].Avian Pathol,1993,22 (2):389-393.
[163]徐超,程安春,汪铭书,等.间接酶免疫组化检测DPV在感染鸭体内细胞定位的研究和应用[J].中国兽医学报,2007,27(5)：640-644.
[164]张小飞,吴亦伦,赵瑞宏,等.免疫酶组化法对试验感染雏鸭体内鸭肝炎病毒的分布测定[J].中国兽医杂志,2005,41(12)：13-15.
[165]陈海军,程安春,汪铭书.Ⅰ型鸭肝炎病毒间接免疫酶染色检测方法的建立[J].中国兽医科学,2007,37(5)：369-373.
[166]Pantin-Jackwood, M.J. Immunohistochemical staining for the detection of the avian influenza virus in tissues[J]. Methods Mol Biol,2008,436:77-83.
[167]Jones, R.C.,Islam, M.R.,Kelly, D.F. Early pathogenesis of experimental reovirus infection in chickens[J].Avian Pathol.1989,18 (2):239-253.
[168]唐雨德,周宗安,翟春生,等.应用免疫酶染色法检测鸡病毒性关节炎病毒的研究[J].中国畜禽传染病,1995,(1)：16-18.
[169]Pertile, T.L.,Walser, M.M.,Sharma, J.M.et al. Immunohistochemical detection of lymphocyte subpopulations in the tarsal joints of chickens with experimental viral arthritis[J]. Vet Pathol,1996,33 (3):303-310.
[170]Fan, Y.H.,Lin, C.D.,Chiou, S.H.et al. Differential expression of U2AF35 in the arthritic joint of avian reovirus-infected chicks[J]. Vet Immunol Immunopathol,2006,114 (1-2):49-60.
[171]Pantin-Jackwood, M.J.,Spackman, E.,Day, J.M. Pathology and virus tissue distribution of turkey origin reoviruses in experimentally infected turkey poults[J]. Vet Pathol,2007,44 (2):185-195.
[172]Shivaprasad, H.L.,Franca, M.,Woolcock, P.R.et al. Myocarditis associated with reovirus in turkey poults[J].Avian Dis,2009,53 (4):523-532.
[173]Sanchez-Cordon, P.J.,Hervas, J.,Chacon de Lara, F.et al. Reovirus infection in psittacine birds (psittacus erithacus):Morphologic and immunohistochemical study[J]. Avian Dis,2002,46 (2):485-492.
[174]Van de Zande, S.,Kuhn, E.M. Central nervous system signs in chickens caused by a new avian reovirus strain:A pathogenesis study[J]. Vet Microbiol,2007,120 (1-2):42-49.
[175]包汉勋.免疫组化法检测番鸭呼肠孤病毒在番鸭体内的分布[D].福州：福建农林大学,2008.
[176]马志永,简中友,姜平,等.斑点酶联免疫吸附试验检测猪繁殖与呼吸综合征病毒抗体方法的建立[J].中国兽医科学,1997,27(4)：3-5.
[177]Albina, E.,Leforban, Y.,Baron, T.et al. An enzyme linked immunosorbent assay (elisa) for the detection of antibodies to the porcine reproductive and respiratory syndrome (prrs) virus[J]. Ann Rech Vet,1992,23 (2):167-176.
[178]蔡雪晖,郭宝清,柴文君,等.检测猪繁殖与呼吸综合征病毒抗体ELISA方法的建立[J].中国预防兽医学报,2000,22(2)：122-125.
[179]Yahara, Y.,Ohkubo, Y.,Kariwa, H.et al. Evaluation of enzyme-linked immunosorbent assay (elisa) and immunofluorescent antibody (IFA) test for the detection of porcine reproductive and respiratory syndrome virus (PRRSV) antibody in pigs from conventional farms[J]. J Vet Med Sci,2002,64 (7):583-588.
[180]Houben, S.,Callebaut, P.,Pensaert, M.B. Comparative study of a blocking enzyme-linked immunosorbent assay and the immunoperoxidase monolayer assay for the detection of antibodies to the porcine reproductive and respiratory syndrome virus in pigs[J]. J Virol Methods,1995,51 (1):125-128.
[181]Ferrin, N.H.,Fang, Y.Johnson, C.R.et al. Validation of a blocking enzyme-linked immunosorbent assay for detection of antibodies against porcine reproductive and respiratory syndrome virus[J]. Clin Diagn Lab Immunol,2004,11(3):503-514.
[182]孙淑芳,包振民,郭福生,等.竞争ELISA检测猪呼吸与繁殖综合征病毒抗体研究[J].中国动物检疫,2006,23(8)：19-22.
[183]de Beer, M.,Peenze, I.,da Costa Mendes, V.M.et al. Comparison of electron microscopy, enzyme-linked immunosorbent assay and latex agglutination for the detection of bovine rotavirus in faeces[J]. J S A fr Vet Assoc,1997,68 (3):93-96.
[184]Schoenthaler, S.L.,Kapil, S. Development and applications of a bovine coronavirus antigen detection enzyme-linked immunosorbent assay[J]. Clin Diagn Lab Immunol,1999,6 (1):130-132.
[185]Monti, G.E.,Frankena, K.,Engel, B.et al. Evaluation of a new antibody-based enzyme-linked immunosorbent assay for the detection of bovine leukemia virus infection in dairy cattle [J]. J Vet Diagn Invest,2005,17 (5):451-457.
[186]Herring, A.J.,Nettleton, P.F.,Burrells, C. A micro-enzyme-linked immunosorbent assay for the detection of antibodies to infectious bovine rhinotracheitis virus[J]. Vet Rec,1980,107 (7):155-156.
[187]杨明凡,崔保安,张素梅,等.鸡传染性喉气管炎病毒gD基因的表达及间接ELISA检测方法的初步建立[J].中国兽医学报,2009,29(3)：263-266.
[188]Jia, R.,Cheng, A.,Wang, M.et al. Development and evaluation of an antigen-capture elisa for detection of the u124 antigen of the duck enteritis virus, based on a polyclonal antibody against the u124 expression protein[J].J Virol Methods,2009,161 (1):38-43.
[189]Zhao, X.L.,Phillips, R.M.,Li, GD.et al. Studies on the detection of antibody to duck hepatitis virus by enzyme-linked immunosorbent assay[J]. Avian Dis,1991,35 (4):778-782.
[190]Wang, M.Y.,Hu, H.L.,Suen, S.Y.et al. Development of an enzyme-linked immunosorbent assay for detecting infectious bursal disease virus (IBDV) infection based on the Vp3 structural protein[J].Vet Microbiol,2008,131 (3-4):229-236.
[191]Slaght, S.S.,Yang, TJ.,van der Heide, L.et al. An enzyme-linked immunosorbent assay (elisa) for detecting chicken anti-reovirus antibody at high sensitivity [J]. Avian Dis,1978,22 (4):802-805.
[192]陈士友,陈溥言.ELISA检测禽呼肠孤病毒抗体方法的建立及应用[J].中国兽医杂 志,1991,17(2)：2-3.
[193]王全溪,吴异健,陈枝华,等.番鸭呼肠病毒抗体间接ELISA方法的建立[J].畜牧与兽医,2005,26(8)：44-47.
[194]Liu, H.J.,Kuo, L.C.,Hu, Y.C.et al. Development of an elisa for detection of antibodies to avian reovirus in chickens[J].J Virol Methods,2002,102 (1-2):129-138.
[195]Yang, ZJ.,Wang, C.Y.,Lee, L.H.et al. Development of elisa kits for antibodies against avian reovirus using the sigmac and sigmab proteins expressed in the methyltropic yeast pichia pastoris[J]. J Virol Methods,2010,163 (2):169-174.
[196]Xie, Z.,Qin, C.,Xie, L.et al. Recombinant protein-based elisa for detection and differentiation of antibodies against avian reovirus in vaccinated and non-vaccinated chickens[J]. J Virol Methods,2010,165(1):108-111.
[197]Zhang, Y.,Guo, D.,Liu, M.et al. Characterization of the sigmab-encoding genes of muscovy duck reovirus:Sigmac-sigmab-elisa for antibodies against duck reovirus in ducks[J]. Vet Microbiol,2007,121 (3-4):231-241.
[198]耿宏伟,郭东春,张云,等.以番鸭呼肠孤病毒σC表达蛋白为抗原的ELISA检测方法的建立[J].中国兽医科学,2006,36(3)：171-176.
[199]谢芝勋,秦宇,谢丽基,等.应用禽呼肠孤病毒σ3融合蛋白为抗原的ELISA检测方法的建立[J].畜牧与兽医,2007,39(11)：3-7.
[200]谢芝勋,秦春香,谢丽基.禽呼肠孤病毒p17蛋白基因在大肠杆菌中的表达及其ELISA检测方法的建立[J].中国兽医科学,2007,37(9)：777-782.
[201]Lin, Y.L.,Shen, J.H.,Lee, L.H. A monoclonal antibody-based competitive enzyme-linked immunosorbent assay for detecting antibody production against avian reovirus protein sigmaa[J]. J Virol Methods,2006,136 (1-2):71-77.
[202]唐雨德,翟春生,周宗安,等.应用ELISA双抗体夹心法检测鸡病毒性关节炎病毒的研究[J].兽医大学学报,1993,13(3)：222-224.
[203]曲立新,幸桂香.用双抗体夹心ELISA法检测鸡病毒性关节炎病毒的研究[J].中国畜禽传染病,1996,(1)：44-46.
[204]沈美艳,张广斌,李久庆,等.呼肠孤病毒在鸡传染性腺胃炎病例中存在情况的研究[J].家畜生态学报,2006,27(6)：112-115.
[205]Leibovitz, L. Gross and histopathological changes of duck plague(duck virus enteritis)[J]. American Journal of veterinary research,1971,32:275-290.
[206]Proctor, S.J.,Matthews, P.J.,Kluge, J.P. Duck plague in gnotobiotic ducks[J]. J Wildl Dis,1976,12 (4):539-544.
[207]Weingarten, M. [duck plague:Clinical aspects, diagnosis, control][J]. Tierarztl Prax,1989,17 (1):53-56.
[208]王伯沄,李玉松,黄高昇.病理学技术[M].人民卫生出版社,2001.
[209]陈晓燕.番鸭呼肠孤病毒感染番鸭的组织学病变及细胞凋亡研究[D].福州：福建农林大学,2007.
[210]刘芳.猪繁殖与呼吸综合征病毒人工感染仔猪的组织超微结构和凋亡的研究[D].雅安：四川农业大学,2005.
[211]Ray, K.,Chakraborty, A.K.,Bhowmik, M.K. Haematological, pathological and certain biochemical changes in experimentally infected indigenous with duck plague virus [J]. Indian Journal of Animal Sciences,1983,53:145-149.
[212]Proctor, S.J. Pathogenesis of digestive tract lesions in duck plague[J]. Vet Pathol,1975,12 (5-6) 349-361.
[213]kaszanyitzky, E.J.,Tanyi, J. Studies on the laboratory diagnosis and epizootiology of duck virus hepatitis[J]. Magy Allatorv Lapja,1980,35:808-814.
[214]Friend, M.,Trainer, D.O. Experimental duck virus hepatitis in the mallard[J]. Avian Dis,1972,16 (4):692-699.
[215]Sturm-Ramirez, K.M.,Ellis, T.,Bousfield, B.et al. Reemerging H5N1 influenza viruses in hong kong in 2002 are highly pathogenic to ducks[J]. J Virol.2004,78 (9):4892-4901.
[216]Cooley, A.J.,Van Campen, H.,Philpott, M.S.et al. Pathological lesions in the lungs of ducks infected with influenza a viruses[J]. Vet Pathol,1989,26 (1):1-5.
[217]Zapata, A.G.,Cooper, E.L. The immune system Comparative histophysiology[M]. Micheser:John Wiley and Sona,1991.
[218]Duncan, R. The low pH-dependent entry of avian reovirus is accompanied by two specific cleavages of the major outer capsid protein & mu;2c[J]. Virology,1996,219 (1):179-189.
[219]Labrada, L.,Bodelon, G.,Vinuela, J.et al. Avian reoviruses cause apoptosis in cultured cells: Viral uncoating, but not viral gene expression, is required for apoptosis induction[J]. J Virol,2002,76 (16):7932-7941.
[220]殷震,刘景华.动物病毒学[M].第二版.北京：科学出版社,1997.
[221]赵亚力,孙衍庆,周捷.外源3型呼肠孤病毒污染原代地鼠肾细胞的光景及电镜诊断[J].中国生物制品学杂志,2001,14(1)：33-35.
[222]Touris-Otero, F.,Cortez-San Martin, M.,Martinez-Costas, J.et al. Avian reovirus morphogenesis occurs within viral factories and begins with the selective recruitment of sigmans and lambdaa to microns inclusions[J]. J Mol Biol,2004,341 (2):361-374.
[223]Touris-Otero, F.,Martinez-Costas, J.,Vakharia, V.N.et al. Avian reovirus nonstructural protein microns forms viroplasm-like inclusions and recruits protein sigmans to these structures [J]. Viro]ogy,2004,319 (1):94-106.
[224]Xu, W.,Patrick, M.K.,Hazelton, P.R.et al. Avian reovirus temperature-sensitive mutant tsal2 has a lesion in major core protein sigmaa and is defective in assembly [J]. J Virol,2004,78 (20):11142-11151.
[225]黄灿华,张建红,高玮.应用光景和电镜对病虾组织细胞病理变化的观察与分析[J].中国病毒学,1997,12(4)：364-370.
[226]Alberts, E. Endoplasmic reticulum:Structure and function molecular biology of the cell[M]. Garland:Garland publishing,1994.
[227]Green, D.E. Mitochondria-structure, function, and replication[J]. N Engl J Med,1983,309 (3):182-183.
[228]Griffiths, E.J. Mitochondria-potential role in cell life and death[J]. Cardiovasc Res,2000,46 (1):24-27.
[229]Bielefeldt Ohmann, H.,Bloch, B. Electron microscopic studies of bovine viral diarrhea virus in tissues of diseased calves and in cell cultures[J]. Arch Virol,1982,71 (1):57-74.
[230]Duncan, R.,Chung, K.L.,Faulkner, P. Analysis of a mutant of autographa California nuclear polyhedrosis virus with a defect in the morphogenesis of the occlusion body macromolecular lattice[J]. J Gen Virol,1983,64 (Pt 7) (1531-1542.
[231]Young, L.S.,Dawson, C.W.,Eliopoulos, A.G. Viruses and apoptosis[J]. Br Med Bull,1997,53 (3):509-521.
[232]Koyama, A.H.,Irie, H.,Fukumori, T.et al. Role of virus-induced apoptosis in a host defense mechanism against virus infection[J]. J Med Invest,1998,45 (1-4):37-45.
[233]Teodoro, J.G.,Branton, P.E. Regulation of apoptosis by viral gene products[J]. J Virol,1997,71 (3):1739-1746.
[234]Clarke, P.,Tyler, K.L. Reovirus-induced apoptosis:A minireview[J]. Apoptosis,2003,8 (2):141-150.
[235]Clarke, P.,Debiasi, R.L.,Goody, R.et al. Mechanisms of reovirus-induced cell death and tissue injury:Role of apoptosis and virus-induced perturbation of host-cell signaling and transcription factor activation[J]. Viral Immunol,2005,18 (1):89-115.
[236]Chulu, J.L.,Lee, L.H.,Lee, Y.C.et al. Apoptosis induction by avian reovirus through p53 and mitochondria-mediated pathway[J]. Biochem Biophys Res Commun,2007,356 (3):529-535.
[237]Lin, H.Y.,Chuang, S.T.,Chen, Y.T.et al. Avian reovirus-induced apoptosis related to tissue injury[J].Avian Pathol,2007,36 (2):155-159.
[238]Kerr, J.F.,Wyllie, A.H.,Currie, A.R. Apoptosis:A basic biological phenomenon with wide-ranging implications in tissue kinetics[J]. Br J Cancer,1972,26 (4):239-257.
[239]Fadeel, B.,Orrenius, S. Apoptosis:A basic biological phenomenon with wide-ranging implications in human disease[J]. J Intern Med,2005,258 (6):479-517.
[240]Oppenheim, R.W. Cell death during development of the nervous system[J]. Annu Rev Neurosci,1991,14:453-501.
[241]Koyama, A.H.,Arakawa, T.,Adachi, A. Characterization of apoptosis induced by sorbitol:A unique system for the detection of antiapoptotic activities of viruses[J]. Microbes Infect,2000,2 (6):599-606.
[242]Miller, L.C.,Fox, J.M. Apoptosis and porcine reproductive and respiratory syndrome virus[J]. Vet Immunol Immunopathol,2004,102 (3):131-142.
[243]Wyllie, A.H.,Kerr, J.F.,Currie, A.R. Cell death:The significance of apoptosis[J]. Int Rev Cytol,1980,68:251-306.
[244]Kerr J.F.R., W.C.M., Harmon B.V.. Apoptosis. Its significance in cancer and cancer therapy [J]. Cancer,1994,73:2013-2026.
[245]Oberhammer F.A., H.K., Fr6sehl G, et al.. Chromatin condensation during apoptosis is accompanied by degradation of lamin a+b, without enhanced activation of cdc2 kinasefJ]. The Journal of Cell Biology,1994,126 (4):827-837.
[246]Strasser, A.,O'Connor, L.,Dixit, V.M. Apoptosis signaling[J]. Annu Rev Biochem,2000,69: 217-245.
[247]Mangili, F.,Cigala, C.,Santambrogio, G. Staining apoptosis in paraffin sections. Advantages and limits[J]. Anal Quant Cytol Histol,1999,21 (3):273-276.
[248]Saraste, A. Morphologic criteria and detection of apoptosis[J]. Herz,1999,24 (3):189-195.
[249]Rihova, Z.,Sefc, L.,Necvas, E. [methods used in the detection of apoptosis][J]. Cas Lek Cesk,2001,140 (6):168-172.
[250]Vaux, D.L.,Haecker, G.,Strasser, A. An evolutionary perspective on apoptosis[J]. Cell,1994,76 (5):777-779.
[251]Jaworowski, A.,Crowe, S.M. Does hiv cause depletion of CD4+t cells in vivo by the induction of apoptosis?[J]. Immunol Cell Biol,1999,77 (1):90-98.
[252]Ito, T.,Kobayashi, Y.,Morita, T.et al. Virulent influenza a viruses induce apoptosis in chickens[J]. Virus Res,2002,84 (1-2):27-35.
[253]Jeurissen, S.H.,Wagenaar, F.,Pol, J.M.et al. Chicken anemia virus causes apoptosis of thymocytes after in vivo infection and of cell lines after in vitro infection[J]. J Virol,1992,66 (12):7383-7388.
[254]Rosenberger, J.K.,Sterner, F.J.,Botts, S.et al. In vitro and in vivo characterization of avian reoviruses. I. Pathogenicity and antigenic relatedness of several avian reovirus isolates[J]. Avian Dis,1989,33 (3):535-544.
[255]DeBiasi, R.L.,Robinson, B.A.,Sherry, B.et al. Caspase inhibition protects against reovirus-induced myocardial injury in vitro and in vivo[J]. J Virol,2004,78 (20):11040-11050.
[256]Salako, M.A.,Carter, M.J.,Kass, G.E. Coxsackievirus protein 2bc blocks host cell apoptosis by inhibiting caspase-3[J]. J Biol Chem,2006,281 (24):16296-16304.
[257]Everett, H.,McFadden, G. Apoptosis:An innate immune response to virus infection[J]. Trends Microbiol,1999,7 (4):160-165.
[258]Dockrell, D.H.,Badley, A.D.,Villacian, J.S.et al. The expression of fas ligand by macrophages and its upregulation by human immunodeficiency virus infection[J]. J Clin Invest,1998,101 (11):2394-2405.
[259]Sabri, F.,Chiodi, F.,Piret, J.P.et al. Soluble factors released by virus specific activated cytotoxic t-lymphocytes induce apoptotic death of astroglioma cell lines[J]. Brain Pathol,2003,13 (2):165-175.
[260]Kawanishi, M. Epstein-barr virus bhrfl protein protects intestine 407 epithelial cells from apoptosis induced by tumor necrosis factor alpha and anti-fas antibody[J]. J Virol,1997,71 (4):3319-3322.
[261]Hay, S.,Kannourakis, G. A time to kill:Viral manipulation of the cell death program[J]. J Gen Virol,2002,83 (Pt7):1547-1564.
[262]Hetts, S.W. To die or not to die:An overview of apoptosis and its role in disease[J]. JAMA,1998,279 (4):300-307.
[263]Shih, W.L.,Hsu, H.W.,Liao, M.H.et al. Avian reovirus sigmac protein induces apoptosis in cultured cells[J].Virology,2004,321 (1):65-74.
[264]陈志胜,马春全,卢玉葵,等.番鸭呼肠孤病毒诱导雏番鸭免疫器官细胞凋亡的研究[J].中国预防兽医学报,2005,27(6)：457-458.
[265]Zamzami, N.,Marchetti, P.,Castedo, M.et al. Reduction in mitochondrial potential constitutes an early irreversible step of programmed lymphocyte death in vivo[J]. J Exp Med,1995,181 (5):1661-1672.
[266]McConkey, D.J.,Nicotera, P.,Orrenius, S. Signalling and chromatin fragmentation in thymocyte apoptosis[J]. Immunol Rev,1994,142:343-363.
[267]Newmeyer, D.D.,Farschon, D.M.,Reed, J.C. Cell-free apoptosis in xenopus egg extracts: Inhibition by Bcl-2 and requirement for an organelle fraction enriched in mitochondria[J]. Cell,1994,79 (2):353-364.
[268]Lee, C.W. Pathogenesis of infectious bronchitis virus with different routes of inoculation and the effect of in vivo serial passage in nephropathogenicity using cloacal infection[J]. Korean Journal of Veterinary Service,2002,25 (1):87-96.
[269]Psalla, D.,Psychas, V.,Spyrou, Vet al. Pathogenesis of experimental encephalomyocarditis: A histopathological, immunohistochemical and virological study in mice[J]. J Comp Pathol,2006,135 (2-3):142-145.
[270]许益民.免疫过氧化物酶染色方法[M].北京：中国农业出版社,2001.
[271]汪家政,范明.蛋白质技术手册[M].北京：科学出版社,2000.
[272]Temponi M., K.T., Perosa F., et al. Purification of murine igg monoclonal antibodies by precipitation with caprylic acid:Comparison with other methods of purification[J]. Hybridoma,1989,8 (1):85-95.
[273]McGuire J.M., D.M., Smith K.D.. The resolution of the neutral n-linked oligosaccharides of igg by high ph anion-exchange chromatography[J]. Carbohydr. Res.,1996,292 (1-9.
[274]John, E.C.精编蛋白质科学实验指南[M].北京：科学出版社,2007.
[275]Sambrook, J., Russell, D. W. Molecular cloning:A. Laboratory manual. [M]. New York:Cold Spring Harbor:Cold Spring Harbor Laboratory Press,2001.
[276]Harlow E.抗体技术实验指南[M].北京：科学出版社,2002.
[277]蔡文琴,王伯法.实用免疫细胞化学与核酸分子杂交技术[M].成都：四川科学技术出版社,1994.
[278]陈立功,刘向平,武现军,等.免疫组化法检测鹅源新城疫病毒在人工感染鹅体内的动态分布[J].中国兽医杂志,2009,(9)：15-17.
[279]Ramos-Vara, J.A.,Beissenherz, M.E. Optimization of immunohistochemical methods using two different antigen retrieval methods on formalin-fixed paraffin-embedded tissues:Experience with 63 markers[J]. J Vet Diagn Invest,2000,12 (4):307-311.
[280]Shi, S.R.,Key, M.E.,Kalra, K.L. Antigen retrieval in formalin-fixed, paraffin-embedded tissues:An enhancement method for immunohistochemical staining based on microwave oven heating of tissue sections[J]. J Histochem Cytochem,1991,39 (6):741-748.
[281]Kahveci, Z.,Minbay, F.Z.,Noyan, S.et al. A comparison of microwave heating and proteolytic pretreatment antigen retrieval techniques in formalin fixed, paraffin embedded tissues[J]. Biotech Histochem,2003,78 (2):119-128.
[282]Hoetelmans, R.W.,van Slooten, HJ.,Keijzer, R.et al. Comparison of the effects of microwave heating and high pressure cooking for antigen retrieval of human and rat be 1-2 protein in formaldehyde-fixed, paraffin-embedded sections[J]. Biotech Histochem,2002,77 (3):137-144.
[283]Gao, C.,Wang, A.Y.,Han, Y.J. Microwave antigen retrieval blocks endogenous peroxidase activity in immunohistochemistry[J]. Appl Immunohistochem Mol Morphol,2008,16 (4):393-399.
[284]Evke, E.,Minbay, F.Z.,Temel, S.Get al. Immunohistochemical detection of p53 protein in basal cell skin cancer after microwave-assisted antigen retrieval [J]. J Mol Histol,2009,40 (1):13-21.
[285]Shi, S.R.,Cote, R.J.,Taylor, C.R. Antigen retrieval immunohistochemistry:Past, present, and future[J].J Histochem Cytochem,1997,45 (3):327-343.
[286]Ramos-Vara, J.A. Technical aspects of immunohistochemistry [J]. Vet Pathol,2005,42 (4):405-426.
[287]Shawky, S. Target cells for duck enteritis virus in lymphoid organs[J]. Avian Pathol,2000,29(6)：609-616.
[288]程安春,韩晓英,汪铭书,等.用间接免疫荧光染色法检测鸭病毒性肠炎病毒在人工感染鸭体内的侵染过程和分布规律[J].畜牧兽医学报,2007,38(9)：942-946.
[289]程国富,胡薛英,周诗其,等.单克隆抗体PAP法对实验感染雏鸭体内鸭肝炎病毒的定位检测[J].华中农业大学学报,1996,15(6)：568-572.
[290]汪铭书,程安春,陈孝跃.鸭病毒性肝炎研究一强毒在雏鸭和成年鸭体内的分布和排泄[J].中国兽医学报,1997,17(3)：254-257.
[291]郑林英.间接夹心ELISA检测鸭肿头败血症病毒的研究[D].雅安：四川农业大学2005.
[292]陈元坤.鸡病毒性关节炎病毒增殖特性与间接ELISA抗体检测方法的建立研究[D].雅安：四川农业大学,2007.
[293]朱立平,陈学清.免疫学常用实验方法[M].北京：人民医学出版社,2000.
[294]McLaren, M.L.,Lillywhite, J.E.,Au, A.C. Indirect enzyme linked immunosorbent assay (ELISA):Practical aspects of standardization and quality control[J]. Med Lab Sci,1981,38 (3):245-251.
[295]Desquesnes, M. [international and regional standardization of immunoenzyme tests: Methods, concerns and limitations][J]. Rev Sci Tech,1997,16 (3):809-823.
[296]Hansson, A.S.,Heinegard, D.,Piette, J.C.et al. The occurrence of autoantibodies to matrilin 1 reflects a tissue-specific response to cartilage of the respiratory tract in patients with relapsing polychondritis[J]. Arthritis Rheum,2001,44 (10):2402-2412.
[297]Lim, K.J.,Park, J.W.,Sohn, M.J.et al. A direct sandwich elisa to detect antibodies against the c-terminal region of merozoite surface protein 1 could be a useful diagnostic method to identify plasmodium vivax exposed persons[J]. Parasitol Res,2002,88 (9):855-860.
[298]Kishimoto, T.,Ando, S.,Numazaki, K.et al. Assay of chlamydia pneumoniae-specific igm antibodies by elisa method-reduction of non-specific reaction and resetting of serological criteria by measuring igm antibodies[J]. Jpn J Infect Dis,2009,62 (4):260-264.
[2]苏敬良,黄瑜.新型鸭肝炎病毒的分离及初步鉴定[J].中国兽医科学,2002,32(1)：15-16.
[3]何冉娅,罗玉均,张桂红,等.一株新型鸭肝炎病毒的分离与初步鉴定[J].养禽与禽病防治,2008,(11)：14-16.
[4]范书才,李虹,袁率珍,等.新型鸭肝炎病毒的分离鉴定[J].中国预防兽医学报,2009,(10):770-775.
[5]Tseng, C.H.,Tsai, H.J. Molecular characterization of a new serotype of duck hepatitis virus[J]. Virus Res,2007,126 (1-2):19-31.
[6]Kim, M.C.,Kwon, Y.K.,Joh, S.J.et al. Recent korean isolates of duck hepatitis virus reveal the presence of a new geno-and serotype when compared to duck hepatitis virus type 1 type strains[J]. Arch Virol,2007,152 (11):2059-2072.
[7]刁有祥,吕桂霞,郑福英,等.一种新型鸭瘟病原的分离鉴定及其特性[J].中国兽医学报,2006,26(2)：136-139.
[8]Vickers, M.L.,Hanson, R.P. Newcastle disease virus in waterfowl in wisconsin[J]. J Wildl Dis,1982,18 (2):149-158.
[9]Spalatin, J.,Hanson, R.P.Epizootiogy of newcastle disease in waterfow[J]. Avian Dis,1975,19 (3):573-582.
[10]张训海,赵家龙.鸭副粘病毒强毒株的分离和鉴定[J].中国动物检疫,2001,18(10)：24-26.
[11]Liu, H.,Wang, Z.,Wang, Y.et al. Characterization of newcastle disease virus isolated from waterfowl in china[J]. Avian Dis,2008,52 (1):150-155.
[12]张太翔,田国宁,张金玲,等.鸭新城疫病毒强毒株的分离和初步鉴定[J].中国兽医杂志,2009,45(9)：35-37.
[13]Sandhu, T.S.,Calnek, B.W.,Zeman, L. Pathologic and serologic characterization of a variant of duck hepatitis type i virus[J]. Avian Dis,1992,36 (4):932-936.
[14]郑献进,张大丙,曲丰发,等.Ⅰ型鸭肝炎病毒变异株的鉴定[J].中国兽医杂志,2006,42(5)：15-16.
[15]柴同杰.山东省樱桃谷鸭群鸭圆环病毒及其混合感染的调查[J].中国家禽,2009,31(6)：4-7.
[16]Soike, D.,Albrecht, K.,Hattermann, K.et al. Novel circovirus in mulard ducks with developmental and feathering disorders[J]. Vet Rec,2004,154 (25):792-793.
[17]吴宝成,陈家祥,姚金水,等.番鸭呼肠孤病毒的分离与鉴定[J].福建农林大学学报(自然科学版),2001,30(2)：227-230.
[18]陈少莺,陈仕龙,林锋强,等.一种新的鸭病(暂名鸭出血性坏死性肝炎)病原学研究初报[J].中国农学通报,2009,25(16)：28-31.
[19]程安春,汪铭书,陈孝跃,等.一种新发现的鸭病毒性肿头出血症的研究[J].中国兽医科学,2003,33(10)：15-21.
[20]张舍郁,程安春,汪铭书,等.间接免疫荧光检测石蜡切片中鸭肿头出血症病毒及抗原定位方法的初步建立[J].中国农业科学,2008,41(3)：868-874.
[21]岳华,罗薇,汤承,等.鸭“传染性肿头症”的研究[J].四川畜牧兽医,2002,29(8)：24-25.
[22]盛敏.鸭肿头败血症病原特性研究[D].雅安：四川农业大学,2003.
[23]Kaleta, E.F. Herpesviruses of birds--areview[J]. Avian Pathol,1990,19 (2):193-211.
[24]Fauquet, C.M.,Mayo, M.A.,Maniloff, J. Virus taxonomy.Ⅷth report of the international committee on taxonomy of virus[M]. San Diego:Elsevier Academic Press,2005.
[25]Nii, S.,Uno, F.,Yoshida, M.et al. [structure and assembly of human beta herpesviruses][J]. Nippon Rinsho,1998,56 (1):22-28.
[26]Kocan, R.M. Duck plague virus replication in muscovy duck fibroblast cells[J]. Avian Dis,1976,20 (3):574-580.
[27]Wolf, K.,Burke, C.N.,Quimby, M.C. Duck viral enteritis:A comparison of replication by ccl-141 and primary cultures of duck embryo fibroblasts[J]. Avian Dis,1976,20 (3):447-454.
[28]Mettenleiter, T.C. Herpesvirus assembly and egress[J]. J Virol,2002,76 (4):1537-1547.
[29]Mettenleiter, T.C.,Klupp, B.G.,Granzow, H. Herpesvirus assembly:A tale of two membranes[J]. Curr Opin Microbiol,2006,9 (4):423-429.
[30]袁桂萍,程安春,汪铭书,等.鸭病毒性肠炎病毒强毒株的形态发生学与超微病理学研究[J].病毒学报,2004,20(4)：326-332.
[31]郭宇飞,程安春,汪铭书,等.鸭病毒性肠炎病毒CH强毒株在鸭胚成纤维细胞中形态发生学的研究[J]．畜牧兽医学报,2006,37(3)：274-280.
[32]Montali, R.J.,Bush, M.,Greenwell, G.A. An epornitic of duck viral enteritis in a zoological park[J].J Am Vet Med Assoc,1976,169 (9):954-958.
[33]Barr, B.C.,Jessup, D.A.,Docherty, D.E.et al. Epithelial intracytoplasmic herpes viral inclusions associated with an outbreak of duck virus enteritis[J]. Avian Dis,1992,36 (1):164-168.
[34]Davison, S.,Converse, K.A.,Hamir, A.N.et al. Duck viral enteritis in domestic muscovy ducks in pennsylvania[J]. Avian Dis,1993,37 (4):1142-1146.
[35]Campagnolo, E.R.,Banerjee, M.,Panigrahy, B.et al. An outbreak of duck viral enteritis (duck plague) in domestic muscovy ducks (cairina moschata domesticus) in illinois[J]. Avian Dis,2001,45 (2):522-528.
[36]彭广能,郭万柱,程安春,等.人工感染鸭瘟的病理学研究[J].中国兽医学报,2009,29(4)：445-450.
[37]Asplin, F.D. Duck hepatitis:Vaccination against two serological types[J]. Vet Rec,1965,77 (50):1529-1530.
[38]Haider, S.A.,Calnek, B.W. In vitro isolation, propagation, and characterization of duck hepatitis virus type iii[J]. Avian Dis,1979,23 (3):715-729.
[39]Tauraso, N.M.,Coghill, G.E.,Klutch, M.J. Properties of the attenuated vaccine strain of duck hepatitis virus[J].Avian Dis,1969,13 (2):321-329.
[40]Kim, M.C.,Kwon, Y.K.,Joh, S J.et al. Molecular analysis of duck hepatitis virus type 1 reveals a novel lineage close to the genus parechovirus in the family picornaviridae[J]. J Gen Virol,2006,87 (Pt 11):3307-3316.
[41]Tseng, C.H.,Knowles, N.J.,Tsai, H.J. Molecular analysis of duck hepatitis virus type 1 indicates that it should be assigned to a new genus[J]. Virus Res,2007,123 (2):190-203.
[42]Ding, C.,Zhang, D. Molecular analysis of duck hepatitis virus type 1[J]. Virology,2007,361 (1):9-17.
[43]Todd, D.,Smyth, V.J.,Ball, N.W.et al. Identification of chicken enterovirus-like viruses, duck hepatitis virus type 2 and duck hepatitis virus type 3 as astroviruses[J]. Avian Pathol,2009,38 (1):21-30.
[44]Toth, T.E. Chicken-embryo-adapted duck hepatitis virus growth curve in embryonated chicken eggs[J]. Avian Dis,1969,13 (3):535-539.
[45]郑献进.鸭肝炎病毒血清型鉴定及Ⅰ型变异株弱毒疫苗的研究[D].北京：中国农业大 学,2006.
[46]Hwang, J. Active immunization against duck hepatitis virus[J]. Am J Vet Res,1972,33 (12):2539-2544.
[47]Davis, D.,Woolcock, P.R. Passage of duck hepatitis virus in cell cultures derived from avian embryos of different species[J]. Res Vet Sci,1986,41 (1):133-134.
[48]Maiboroda, A.D. [cultivation of duck hepatitis virus in cell culture][J]. Veterinariia,1972,8: 50-52.
[49]Doroshko, I.N.,Bezrukavaia, I.,Golubnichii, V.P.et al. [production tests of a vaccine against viral hepatitis in ducks][J]. Veterinariia,1975, (1):52-54.
[50]Richter, W.R.,Rdzok, E.J.,Moize, S.M. Electron microscopy of viruslike particles associated with duck viral hepatitis[J]. Virology,1964,24:114-116.
[51]谢毅,邝荣禄,林维庆.鸭肝炎病毒形态及接种鸡胚引起的病理变化观察[J].华南农业大学学报,1989,10(4)：65-71.
[52]徐福南,周芳.鸭病毒性肝炎的组织病理学研究[J].中国兽医科学,1990,(2)：6-7.
[53]胡薛英,程国富.试验感染鸭病毒性肝炎雏鸭的组织病理学研究[J].华南农业大学学报,2000,19(1)：48-50.
[54]彭南秀.雏鸭病毒性肝炎的动态病理学研究[J].中国兽医科学,1998,28(8)：22-23.
[55]Farmer, H.,Chalmers, W.S.,Woolcock, P.R. The duck fatty kidney syndrome-an aspect of duck viral hepatitis[J]. Avian Pathol,1987,16 (2):227-236.
[56]陈建红,梁发朝,卢玉葵.鸭肝炎自然病例与鸭肝炎病毒人工发病病例组织病理学观察[J].中国预防兽医学报,2001,23(3)：203-205.
[57]祁保民,姚金水,卢惠明.鸭病毒性肝炎的病理形态学观察[J].福建农林大学学报(自然科学版),1999,28(3)：350-352.
[58]胡薛英,谷长勤,王德海,等.雏鸭胰脏组织损伤在新型鸭肝炎病毒感染过程中的变化特点[J].中国兽医学报,2006,26(2)：186-188.
[59]王永坤.禽流感流行新特点及防制对策[J].中国家禽,2002,24(12)：6-8.
[60]Fouchier, R.A.,Munster, V.,Wallensten, A.et al. Characterization of a novel influenza a virus hemagglutinin subtype (h16) obtained from black-headed gulls[J]. J Virol,2005,79 (5):2814-2822.
[61]Booy, F.P.,Ruigrok, R.W.,van Bruggen, E.F. Electron microscopy of influenza virus. A comparison of negatively stained and ice-embedded particles[J]. J Mol Biol,1985,184 (4)：667-676.
[62]田国斌,赵增连,唐秀英,等.鸭源禽流感病毒的分离和鉴定[J].中国预防兽医学报,1999,21(1)：1-3.
[63]李银,刘宇卓,张敬峰,等.一株鸭源H9亚型禽流感病毒在鸡胚上传代及繁殖规律的研究[J].江苏农业科学,2006,(4)：94-96.
[64]Briedis, D.J.,Conti, G.,Munn, E.A.et al. Migration of influenza virus-specific polypeptides from cytoplasm to nucleus of infected cells[J]. Virology,1981,111(1):154-164.
[65]Choppin, P.W.,Scheid, A. The role of viral glycoproteins in adsorption, penetration, and pathogenicity of viruses[J]. Rev Infect Dis,1980,2 (1):40-61.
[66]Krug, R.M.,Soeiro, R. Studies on the intranuclear localization of influenza virus-specific proteins[J].Virology,1975,64 (2):378-387.
[67]Compans, R.W.,Meier-Ewert, H.,Palese, P. Assembly of lipid-containing viruses[J]. J Supramol Struct,1974,2 (2-4):496-511.
[68]Franklin, R.M.,Hinnen, R.,Schafer, R.et al. Structure and assembly of lipid-containing viruses, with special reference to bacteriophage pm2 as one type of model system[J]. Philos Trans R Soc Lond B Biol Sci,1976,276 (943):63-80.
[69]李银.禽流感病毒H9N2鸭分离株的生物学与分子生物学特性及其快速诊断的研究[D].南京：南京农业大学,2006.
[70]刘超男,刘明,张云,等.H5N1亚型禽流感病毒的鸭致病性研究[J].中国农业科学,2006,39(2)：412-417.
[71]李玉谷,周泉鹤,崔聪颖,等.一株H5N1亚型高致病力禽流感病毒人工感染鸭的组织病理学观察[J].畜牧兽医学报,2008,39(10)：1373-1381.
[72]马春全,卢玉葵,邓桦,等.鸭H5N1型高致病性禽流感的病理组织学观察[J].中国兽医科学,2004,34(11)：11-14.
[73]岳华,刘群,汤承,等.“鸭传染性肿头症”病毒的分离及部分特性研究[J].西南民族学院学报,2002,28(3)：314-316.
[74]岳华,刘群,汤承,等.”鸭传染性肿头症”病毒对雏鸭致病性和卵黄抗体研究[J].西南民族学院学报,2002,28(4)：519-520.
[75]何平有,蒋文灿.实验性鸭肿头病败血症病理研究[J].黑龙江畜牧兽医,2006,11)：87-88.
[76]张舍郁,朱德康,程安春,等.鸭肿头出血症病毒强毒的致病性及感染组织细胞的超微病 理变化[J].中国兽医科学,2007,37(2)：1017-1023.
[77]张舍郁,程安春,汪铭书.鸭病毒性肿头出血症病毒培养特性与形态观察[J].黑龙江畜牧兽医,2007,(11)：9-11.
[78]杨汉春.动物免疫学[M].第二版.北京：中国农业大学出版社,2003.
[79]郭积燕.免疫学检验中的酶免疫技术[J].中华检验医学杂志,2005,28(2)：221-224.
[80]Voller, A.,Bidwell, D.E.,Bartlett, A. Enzyme immunoassays in diagnostic medicine. Theory and practice[J]. Bull World Health Organ,1976,53 (1):55-65.
[81]Nakane, P.K.,Pierce, G.B., Jr. Enzyme-labeled antibodies:Preparation and application for the localization of antigens[J]. J Histochem Cytochem,1966,14 (12):929-931.
[82]Avrameas, S.,Uriel, J. Method for labeling antigens and antibodies with enzymes and their application in immunodiffusion[J]. Compt Rend SerD,1966,262:2543-2545.
[83]Ram, J., S.,Nakane, P.K.,Rawlinson, D.G.et al. Enzyme-labelled antibodies for ultrastructural studies[J]. Fed Proc,1966,25:732.
[84]Miles, L.E.,Hales, C.N. Labelled antibodies and immunological assay systems[J]. Nature,1968,219 (5150):186-189.
[85]Avrameas, S. Coupling of enzymes to proteins with glutaraldehyde. Use of the conjugates for the detection of antigens and antibodies[J]. Immunochemistry,1969,6 (1):43-52.
[86]Engvall, E.,Perlmann, P. Enzyme-linked immunosorbent assay (ELISA). Quantitative assay of immunoglobuling[J]. Immunochemistry,1971,8 (9):871-874.
[87]Van Weemen, B.K.,Schuurs, A.H. Immunoassay using antigen-enzyme conjugates[J]. FEBS Lett,1971,15:232-235.
[88]Rubenstein, K.E.,Schneider, R.S.,Ullman, E.F. "Homogeneous" enzyme immunoassay. A new immunochemical technique[J]. Biochem Biophys Res Commun,1972,47 (4):846-851.
[89]Matsuoka, K.,Maeda, M.,Tsuji, A. Fluorescence enzyme immunoassay for insulin using peroxidase-tyramine-hydrogen peroxide[J]. Chem Pharm Bull (Tokyo),1979,27 (10):2345-2350.
[90]Wang, H.X.,Stott, R.A.,Thorpe, GH.et al. An enhanced chemiluminescence enzyme immunoassay for serum progesterone[J]. Steroids,1984,44 (4):317-328.
[91]Halsall, H.B.,Heineman, W.R. Electrochemical immunoassay:An ultrasensitive method[J]. J Int Fed Clin Chem,1990,2 (4):179-187.
[92]Alard, P.,Lantz, O.,Sebagh, M.et al. A versatile elisa-pcr assay for mma quantitation from a few cells[J].Biotechniques,1993,15 (4):730-737.
[93]Wu, M.,Stockley, P.G.,Martin, W.J.,2nd. An improved western blotting technique effectively reduces background[J]. Electrophoresis,2002,23 (15):2373-2376.
[94]Ahangaran, A.,Mohammadi, G.M.,Habibi, M.K.et al. Rapid diagnosis of soybean mosaic virus n soybean by tissue-print immunoassay and diba in comparison to other serological methods[J]. Commun Agric Appl Biol Sci,2006,71 (3PtB):1207-1212.
[95]钟晓琴,刘美英,黄云艳,等.一种基于酶标纳米金探针的蛋白质检测方法[J].中国医药生物技术,2009,4(3)：200-206.
[96]Sternberger, L.A. Immunocytochemistry[M].3rd. New York:John Wiley&Sons,1986.
[97]Maekawa, M. [properties of enzymes for enzyme immunoassay][J]. Nippon Rinsho,1995,53 (9):2154-2159.
[98]Krieg, R.,Halbhuber, K.J. Recent advances in catalytic peroxidase histochemistry[J]. Cell Mol Biol (Noisy-le-grand),2003,49 (4):547-563.
[99]Porstmann, T.,Kiessig, S.T. Enzyme immunoassay techniques. An overview[J]. J Immunol Methods,1992,150 (1-2):5-21.
[100]Ormanns, W.,Schaffer, R. An alkaline-phosphatase staining method in avidin-biotin immunohistochemistry[J]. Histochemistry,1985,82 (5):421-424.
[101]尹伯元,王仁芝,李振甲.标记免疫学[M].北京：原子能出版社,1998.
[102]Hancock, M.B. Leu-enkephalin, substance p, and somatostatin immunohistochemistry combined with the retrograde transport of horseradish peroxidase in sympathetic preganglionic neurons[J].J Auton Nerv Syst,1982,6 (2):263-272.
[103]Yamashita, S.,Korach, K.S. A modified immunohistochemical procedure for the detection of estrogen receptor in mouse tissues[J]. Histochemistry,1989,90 (5):325-330.
[104]Erdogan, D.,Elmas, C.,Ilgaz, C. Visualization of sperm by immunohistochemistry: Application of alternative counterstains[J]. Arch Androl,2005,51 (3):233-238.
[105]Chi, V.,Chandy, K.G. Immunohistochemistry:Paraffin sections using the vectastain abc kit from vector labs[J].J Vis Exp,2007, (8):308.
[106]Jiao, K.,Sun, W.,Zhang, S.S. Sensitive detection of a plant virus by electrochemical enzyme-linked immunoassay [J]. Fresenius J Anal Chem,2000,367 (7):667-671.
[107]Zelnik, V.,Harlin, O.,Fehler, F.et al. An enzyme-linked immunosorbent assay (ELISA) for detection of marek's disease virus-specific antibodies and its application in an experimental vaccine trial[J]. J Vet Med B Infect Dis Vet Public Health,2004,51 (2):61-67.
[108]Benbrook, D.M. An elisa method for detection of human antibodies to an immunotoxin[J]. J Pharmacol Toxicol Methods,2002,47 (3):169-175.
[109]Reyna-Bello, A.,Eleizalde, M.C.,Silva, A.M. Assessment of chromogen suitability in elisa for the detection of anaplasmosis and trypanosomosis[J]. J Immunoassay Immunochem,2007,28 (1):1-11.
[110]Khatun, S.,Nara, S.,Tripathi, Vet al. Development of elisa for measurement of progesterone employing 17-alpha-oh-p-hrp as enzyme label[J]. J Immunoassay Immunochem,2009,30 (2):186-196.
[111]Jiao, K.,YAO, H.,Xu, J.et al. Electrochemical studies of 1,5-dihydroxynaphthalene as substrate for voltammetric enzyme immunoassay[J]. Sci China Ser B,2004,47 (3):184-191.
[112]Jiao, G.Z.,Zhu, L.H.,Yao, H.et al. Voltammetric enzyme-linked immunoassay for dectection of horseradish peroxidase using phenol red as substrate [J]. J Qingdao Univ Sci & Tech,2004,25 (2):98-101.
[113]Zhang, S.S.,Jiao, K.,Chen, H.Y. Investigation of voltammetric enzyme-linked immunoassay based on a new system of oap-h2o2-hrp[J]. Electroanalysis,1999,11(7):511-516.
[114]Bobrow, M.N.,Harris, T.D.,Shaughnessy, KJ.et al. Catalyzed reporter deposition, a novel method of signal amplification. Application to immunoassays[J]. J Immunol Methods,1989,125 (1-2):279-285.
[115]Adams, J.C. Biotin amplification of biotin and horseradish peroxidase signals in histochemical stains[J]. J Histochem Cytochem,1992,40 (10):1457-1463.
[116]Mayer, G.,Bendayan, M. Biotinyl-tyramide:A novel approach for electron microscopic immunocytochemistry[J]. J Histochem Cytochem,1997,45 (11):1449-1454.
[117]Kohler, A.,Lauritzen, B.,Van Noorden, C.J. Signal amplification in immunohistochemistry at the light microscopic level using biotinylated tyramide and nanogold-silver staining[J]. J Histochem Cytochem,2000,48 (7):933-941.
[118]Hikino, T. In situ hybridization with novel biotinyl-tyramide:Fundamental studies and its utility of the detection of human papilloma virus in tissue sections[J]. Rinsho Byori,2007,55 (10):922-929.
[119]Krieg, R.,Halbhuber, K.J. Novel oxidative self-anchoring fluorescent substrates for the histochemical localization of endogenous and immunobound peroxidase activity[J]. J Mol Histol,2004,35 (5):471-487.
[120]沈关心,周汝霖.现代免疫学实验技术[M].武汉：湖北科学技术出版社,1998.
[121]Tang, B.,Wang, Y.,Du, M.et al. Catalytic spectrophotometric determination of ascorbic acid in tea drink with 1,5-bis(p-hydroxybenzaldene)thiocarbohydrazone as the substrate for horseradish peroxidase[J].JAgric Food Chem,2003,51 (15):4198-4201.
[122]Ichibangase, T.,Ohba, Y.,Kishikawa, N.et al. Chemiluminescence assay of lipase activity using a synthetic substrate as proenhancer for luminol chemiluminescence reaction[J]. Luminescence,2004,19 (5):259-264.
[123]Hainfeld, J.F. Enzymatic deposition and alteration of metals[P]. U.S. Patent,6670113,2003.
[124]Morgenstern, S.,Kessler, G.,Auerbach, J.et al. An automated p-nitrophenylphosphate serum alkaline phosphatase procedure for the autoanalyzer[J]. Clin Chem,1965,11 (9):876-888.
[125]Neumann, H.,van Vreedendaal, M. An improved alkaline phosphatase determination with p-nitrophenyl phosphate[J].Clin Chim Acta,1967,17 (2):183-187.
[126]Geneix, N.,Dufour, E.,Venien, A.et al. Development of a monoclonal antibody-based immunoassay for specific quantification of bovine milk alkaline phosphatase[J]. J Dairy Res,2007,74 (3):290-295.
[127]Anyaoku, V.N.,Wood, D.F.,Pavlou, R.et al. An enzyme-linked immunoadsorbent assay for human pituitary glycoprotein free alpha-subunit:Evaluation and initial experience in patients with pituitary disease[J].J Endocrinol,1993,136 (3):511-516.
[128]Avrameas, S.,Ternynck, T. Peroxidase labelled antibody and fab conjugates with enhanced intracellular penetration[J]. Immunochemistry,1971,8 (12):1175-1179.
[129]Sternberger, L.A. Immunocytochemistry[M].2nd. New York:John Wiley & Sons,1979.
[130]Imagawa, M.,Yoshitake, S.,Ishikawa, E.et al. Development of a highly sensitive sandwich enzyme immunoassay for human ferritin using affinity-purified anti-ferritin labelled with beta-d-galactosidase from escherichia coli[J]. Clin Chim Acta,1982,121 (3):277-289.
[131]Herrmann, J.E.,Morse, S.A. Coupling of peroxidase to poliovirus antibody:Characteristics of the conjugates and their use in virus detection[J]. Infect Immun,1973,8 (4):645-649.
[132]Wisdom, G.B. Conjugation of antibodies to horseradish peroxidase [J]. Methods Mol Biol,2005,295:127-130.
[133]Petrali, J.P.,Hinton, D.M.,Moriarty, GC.et al. The unlabeled antibody enzyme method of immunocytochemistry. Quantitative comparison of sensitivities with and without peroxidase-antiperoxidase complex[J]. J Histochem Cytochem,1974,22 (8):782-801.
[134]Sinclair, R.A.,Burns, J.,Dunnill, M.S. Immunoperoxidase staining of formalin-fixed, paraffin-embedded, human renal biopsies with a comparison of the peroxidase-antiperoxidase (pap) and indirect methods[J]. J Clin Pathol,1981,34 (8):859-865.
[135]Vacca, L.L.,Rosario, S.L.,Zimmerman, E.A.et al. Application of immunoperoxidase techniques to localize horseradish peroxidase-tracer in the central nervous system[J]. J Histochem Cytochem,1975,23 (3):208-215.
[136]Mason, D.Y.,Abdulaziz, Z.,Falini, B.et al. Single and double immunoenzymatic techniques for labeling tissue sections with monoclonal antibodies [J]. Ann N Y Acad Sci,1983,420:127-133.
[137]Moir, D.J.,Ghosh, A.K.,Abdulaziz, Z.et al. Immunoenzymatic staining of haematological samples with monoclonal antibodies [J]. Br J Haematol,1983,55 (3):395-410.
[138]Chilosi, M.,Lestani, M.,Pedron, S.et al. A rapid immunostaining method for frozen sections[J]. Biotech Histochem,1994,69 (4):235-239.
[139]Sabattini, E.,Bisgaard, K.,Ascani, S.et al. The envision++system:A new immunohistochemical method for diagnostics and research. Critical comparison with the apaap, chemmate, csa, labc, and sabc techniques[J]. J Clin Pathol,1998,51 (7):506-511.
[140]Shi, S.R.,Guo, J.,Cote, R.J.et al. Sensitivity and detection efficiency of a novel two-step detection system (powervision) for immunohistochemistry[J]. Appl. Immunohistochem. Mol. Morphol.,1999,7:201-208.
[141]Ogawa, T.,Gamoh, K.,Aoki, H.et al. Validation and standardization of virus neutralizing test using indirect immunoperoxidase technique for the quantification of antibodies to rabies virus[J]. Zoonoses Public Health,2008,55 (6):323-327.
[142]Hoffman, D.R. Enzyme-linked immunosorbent assays (ELISA) for immunoglobulin e and blocking antibodies[J]. Methods Enzymol,1981,73 (PtB):656-666.
[143]Hawkes, R.,Niday, E.,Gordon, J. A dot-immunobinding assay for monoclonal and other antibodies[J].Anal Biochem,1982,119 (1):142-147.
[144]Herbrink, P.,Van Bussel, F.J.,Warnaar,S.O. The antigen spot test (AST):A highly sensitive assay for the detection of antibodies [J]. J Immunol Methods,1982,48 (3):293-298.
[145]Karawajew, L.,Behrsing, O.,Kaiser, Get al. Production and elisa application of bispecific monoclonal antibodies against fluorescein isothiocyanate (FITC) and horseradish peroxidase (HRP)[J].J Immunol Methods,1988,111(1):95-99.
[146]Schwab, K.J.,McDevitt, J.J. Development of a PCR-enzyme immunoassay oligoprobe detection method for toxoplasma gondii oocysts, incorporating pcr controls [J]. Appl Environ Microbiol,2003,69 (10):5819-5825.
[147]Whiley, D.M.,Arden, K.E.,Mackay, I.M.et al. Simultaneous detection and differentiation of human polyomaviruses jc and bk by a rapid and sensitive PCR-elaha assay and a survey of the jcv subtypes within an australian population[J]. J Med Virol,2004,72 (3):467-472.
[148]Nguyen, T.D.,de Kesel, M.,Bigaignon, Get al. Detection of toxoplasma gondii tachyzoites and bradyzoites in blood, urine, and brains of infected mice[J]. Clin Diagn Lab Immunol,1996,3 (6):635-639.
[149]Zhou, M.,Liu, Q.,Wongsrichanalai, C.et al. High prevalence of plasmodium malariae and plasmodium ovale in malaria patients along the thai-myanmar border, as revealed by acridine orange staining and pcr-based diagnoses[J]. Trop Med Int Health,1998,3 (4):304-312.
[150]Bora, U.,Kannan, K.,Nahar, P. Heat-mediated enzyme-linked immunosorbent assay procedure on a photoactivated surface[J]. J Immunol Methods,2004,293 (1-2):43-50.
[151]Sharma, P.,Nahar, P. Ultrasound wave-mediated enzyme-linked immunosorbent assay technique[J].Anal Chim Acta,2009,650 (2):241-246.
[152]马吉飞,邱震东,郑兆荣,等.应用免疫组化PAP法检测兔出血症病毒抗原的动态分布[J].中国兽医学报,1994,14(3)：264-267.
[153]Halbur, P.G,Miller, L.D.,Paul, P.S.et al. Immunohistochemical identification of porcine reproductive and respiratory syndrome virus (PRRSV) antigen in the heart and lymphoid system of three-week-old colostrum-deprived pigs[J]. Vet Pathol,1995,32 (2):200-204.
[154]Last, R.D.,Jardine, J.E.,Smit, M.M.et al. Application of immunoperoxidase techniques to formalin-fixed brain tissue for the diagnosis of rabies in southern africa[J]. Onderstepoort J Vet Res,1994,61 (2):183-187.
[155]Clift, S.J.,Williams, M.C.,Gerdes, T.et al. Standardization and validation of an immunoperoxidase assay for the detection of african horse sickness virus in formalin-fixed, paraffin-embedded tissues[J]. J Vet Diagn Invest,2009,21(5):655-667.
[156]Carlsson, G.L.,Heller, R.S.,Serup, P.et al. Immunohistochemistry of pancreatic development in cattle and pig[J].Anat Histol Embryol,2010,39 (2):107-119.
[157]张晓梅,杨汉春,任慧英,等.用LSAB免疫组化染色法检测猪生殖-呼吸道综合征病毒抗原的研究[J].中国预防兽医学报,1999,21(6)：427-429.
[158]张平英,朱德康,程安春,等.间接免疫酶组织化学法检测猪繁殖与呼吸综合征病毒感染和抗原定位[J].中国兽医学报,2008,28(2)：128-132.
[159]Haines, D.M.,Moline, K.M.,Sargent, R.A.et al. Immunohistochemical study of hemophilus somnus, mycoplasma bovis, mannheimia hemolytica, and bovine viral diarrhea virus in death losses due to myocarditis in feedlot cattle[J]. Can Vet J,2004,45 (3):231-234.
[160]Njaa, B.L.,Clark, E.G,Janzen, E.et al. Diagnosis of persistent bovine viral diarrhea virus infection by immunohistochemical staining of formalin-fixed skin biopsy specimens[J]. J Vet Diagn Invest,2000,12 (5):393-399.
[161]Dougherty, R.M.,Marucci, A.A.,Distefano, H.S. Application of immunohistochemistry to study of avian leukosis virus[J].J Gen Virol,1972,15 (2):149-162.
[162]Islam, M.R.,Nessa, J.,Halder, K.M. Detection of duck plague virus antigen in tissues by immunoperoxidase staining[J].Avian Pathol,1993,22 (2):389-393.
[163]徐超,程安春,汪铭书,等.间接酶免疫组化检测DPV在感染鸭体内细胞定位的研究和应用[J].中国兽医学报,2007,27(5)：640-644.
[164]张小飞,吴亦伦,赵瑞宏,等.免疫酶组化法对试验感染雏鸭体内鸭肝炎病毒的分布测定[J].中国兽医杂志,2005,41(12)：13-15.
[165]陈海军,程安春,汪铭书.Ⅰ型鸭肝炎病毒间接免疫酶染色检测方法的建立[J].中国兽医科学,2007,37(5)：369-373.
[166]Pantin-Jackwood, M.J. Immunohistochemical staining for the detection of the avian influenza virus in tissues[J]. Methods Mol Biol,2008,436:77-83.
[167]Jones, R.C.,Islam, M.R.,Kelly, D.F. Early pathogenesis of experimental reovirus infection in chickens[J].Avian Pathol.1989,18 (2):239-253.
[168]唐雨德,周宗安,翟春生,等.应用免疫酶染色法检测鸡病毒性关节炎病毒的研究[J].中国畜禽传染病,1995,(1)：16-18.
[169]Pertile, T.L.,Walser, M.M.,Sharma, J.M.et al. Immunohistochemical detection of lymphocyte subpopulations in the tarsal joints of chickens with experimental viral arthritis[J]. Vet Pathol,1996,33 (3):303-310.
[170]Fan, Y.H.,Lin, C.D.,Chiou, S.H.et al. Differential expression of U2AF35 in the arthritic joint of avian reovirus-infected chicks[J]. Vet Immunol Immunopathol,2006,114 (1-2):49-60.
[171]Pantin-Jackwood, M.J.,Spackman, E.,Day, J.M. Pathology and virus tissue distribution of turkey origin reoviruses in experimentally infected turkey poults[J]. Vet Pathol,2007,44 (2):185-195.
[172]Shivaprasad, H.L.,Franca, M.,Woolcock, P.R.et al. Myocarditis associated with reovirus in turkey poults[J].Avian Dis,2009,53 (4):523-532.
[173]Sanchez-Cordon, P.J.,Hervas, J.,Chacon de Lara, F.et al. Reovirus infection in psittacine birds (psittacus erithacus):Morphologic and immunohistochemical study[J]. Avian Dis,2002,46 (2):485-492.
[174]Van de Zande, S.,Kuhn, E.M. Central nervous system signs in chickens caused by a new avian reovirus strain:A pathogenesis study[J]. Vet Microbiol,2007,120 (1-2):42-49.
[175]包汉勋.免疫组化法检测番鸭呼肠孤病毒在番鸭体内的分布[D].福州：福建农林大学,2008.
[176]马志永,简中友,姜平,等.斑点酶联免疫吸附试验检测猪繁殖与呼吸综合征病毒抗体方法的建立[J].中国兽医科学,1997,27(4)：3-5.
[177]Albina, E.,Leforban, Y.,Baron, T.et al. An enzyme linked immunosorbent assay (elisa) for the detection of antibodies to the porcine reproductive and respiratory syndrome (prrs) virus[J]. Ann Rech Vet,1992,23 (2):167-176.
[178]蔡雪晖,郭宝清,柴文君,等.检测猪繁殖与呼吸综合征病毒抗体ELISA方法的建立[J].中国预防兽医学报,2000,22(2)：122-125.
[179]Yahara, Y.,Ohkubo, Y.,Kariwa, H.et al. Evaluation of enzyme-linked immunosorbent assay (elisa) and immunofluorescent antibody (IFA) test for the detection of porcine reproductive and respiratory syndrome virus (PRRSV) antibody in pigs from conventional farms[J]. J Vet Med Sci,2002,64 (7):583-588.
[180]Houben, S.,Callebaut, P.,Pensaert, M.B. Comparative study of a blocking enzyme-linked immunosorbent assay and the immunoperoxidase monolayer assay for the detection of antibodies to the porcine reproductive and respiratory syndrome virus in pigs[J]. J Virol Methods,1995,51 (1):125-128.
[181]Ferrin, N.H.,Fang, Y.Johnson, C.R.et al. Validation of a blocking enzyme-linked immunosorbent assay for detection of antibodies against porcine reproductive and respiratory syndrome virus[J]. Clin Diagn Lab Immunol,2004,11(3):503-514.
[182]孙淑芳,包振民,郭福生,等.竞争ELISA检测猪呼吸与繁殖综合征病毒抗体研究[J].中国动物检疫,2006,23(8)：19-22.
[183]de Beer, M.,Peenze, I.,da Costa Mendes, V.M.et al. Comparison of electron microscopy, enzyme-linked immunosorbent assay and latex agglutination for the detection of bovine rotavirus in faeces[J]. J S A fr Vet Assoc,1997,68 (3):93-96.
[184]Schoenthaler, S.L.,Kapil, S. Development and applications of a bovine coronavirus antigen detection enzyme-linked immunosorbent assay[J]. Clin Diagn Lab Immunol,1999,6 (1):130-132.
[185]Monti, G.E.,Frankena, K.,Engel, B.et al. Evaluation of a new antibody-based enzyme-linked immunosorbent assay for the detection of bovine leukemia virus infection in dairy cattle [J]. J Vet Diagn Invest,2005,17 (5):451-457.
[186]Herring, A.J.,Nettleton, P.F.,Burrells, C. A micro-enzyme-linked immunosorbent assay for the detection of antibodies to infectious bovine rhinotracheitis virus[J]. Vet Rec,1980,107 (7):155-156.
[187]杨明凡,崔保安,张素梅,等.鸡传染性喉气管炎病毒gD基因的表达及间接ELISA检测方法的初步建立[J].中国兽医学报,2009,29(3)：263-266.
[188]Jia, R.,Cheng, A.,Wang, M.et al. Development and evaluation of an antigen-capture elisa for detection of the u124 antigen of the duck enteritis virus, based on a polyclonal antibody against the u124 expression protein[J].J Virol Methods,2009,161 (1):38-43.
[189]Zhao, X.L.,Phillips, R.M.,Li, GD.et al. Studies on the detection of antibody to duck hepatitis virus by enzyme-linked immunosorbent assay[J]. Avian Dis,1991,35 (4):778-782.
[190]Wang, M.Y.,Hu, H.L.,Suen, S.Y.et al. Development of an enzyme-linked immunosorbent assay for detecting infectious bursal disease virus (IBDV) infection based on the Vp3 structural protein[J].Vet Microbiol,2008,131 (3-4):229-236.
[191]Slaght, S.S.,Yang, TJ.,van der Heide, L.et al. An enzyme-linked immunosorbent assay (elisa) for detecting chicken anti-reovirus antibody at high sensitivity [J]. Avian Dis,1978,22 (4):802-805.
[192]陈士友,陈溥言.ELISA检测禽呼肠孤病毒抗体方法的建立及应用[J].中国兽医杂 志,1991,17(2)：2-3.
[193]王全溪,吴异健,陈枝华,等.番鸭呼肠病毒抗体间接ELISA方法的建立[J].畜牧与兽医,2005,26(8)：44-47.
[194]Liu, H.J.,Kuo, L.C.,Hu, Y.C.et al. Development of an elisa for detection of antibodies to avian reovirus in chickens[J].J Virol Methods,2002,102 (1-2):129-138.
[195]Yang, ZJ.,Wang, C.Y.,Lee, L.H.et al. Development of elisa kits for antibodies against avian reovirus using the sigmac and sigmab proteins expressed in the methyltropic yeast pichia pastoris[J]. J Virol Methods,2010,163 (2):169-174.
[196]Xie, Z.,Qin, C.,Xie, L.et al. Recombinant protein-based elisa for detection and differentiation of antibodies against avian reovirus in vaccinated and non-vaccinated chickens[J]. J Virol Methods,2010,165(1):108-111.
[197]Zhang, Y.,Guo, D.,Liu, M.et al. Characterization of the sigmab-encoding genes of muscovy duck reovirus:Sigmac-sigmab-elisa for antibodies against duck reovirus in ducks[J]. Vet Microbiol,2007,121 (3-4):231-241.
[198]耿宏伟,郭东春,张云,等.以番鸭呼肠孤病毒σC表达蛋白为抗原的ELISA检测方法的建立[J].中国兽医科学,2006,36(3)：171-176.
[199]谢芝勋,秦宇,谢丽基,等.应用禽呼肠孤病毒σ3融合蛋白为抗原的ELISA检测方法的建立[J].畜牧与兽医,2007,39(11)：3-7.
[200]谢芝勋,秦春香,谢丽基.禽呼肠孤病毒p17蛋白基因在大肠杆菌中的表达及其ELISA检测方法的建立[J].中国兽医科学,2007,37(9)：777-782.
[201]Lin, Y.L.,Shen, J.H.,Lee, L.H. A monoclonal antibody-based competitive enzyme-linked immunosorbent assay for detecting antibody production against avian reovirus protein sigmaa[J]. J Virol Methods,2006,136 (1-2):71-77.
[202]唐雨德,翟春生,周宗安,等.应用ELISA双抗体夹心法检测鸡病毒性关节炎病毒的研究[J].兽医大学学报,1993,13(3)：222-224.
[203]曲立新,幸桂香.用双抗体夹心ELISA法检测鸡病毒性关节炎病毒的研究[J].中国畜禽传染病,1996,(1)：44-46.
[204]沈美艳,张广斌,李久庆,等.呼肠孤病毒在鸡传染性腺胃炎病例中存在情况的研究[J].家畜生态学报,2006,27(6)：112-115.
[205]Leibovitz, L. Gross and histopathological changes of duck plague(duck virus enteritis)[J]. American Journal of veterinary research,1971,32:275-290.
[206]Proctor, S.J.,Matthews, P.J.,Kluge, J.P. Duck plague in gnotobiotic ducks[J]. J Wildl Dis,1976,12 (4):539-544.
[207]Weingarten, M. [duck plague:Clinical aspects, diagnosis, control][J]. Tierarztl Prax,1989,17 (1):53-56.
[208]王伯沄,李玉松,黄高昇.病理学技术[M].人民卫生出版社,2001.
[209]陈晓燕.番鸭呼肠孤病毒感染番鸭的组织学病变及细胞凋亡研究[D].福州：福建农林大学,2007.
[210]刘芳.猪繁殖与呼吸综合征病毒人工感染仔猪的组织超微结构和凋亡的研究[D].雅安：四川农业大学,2005.
[211]Ray, K.,Chakraborty, A.K.,Bhowmik, M.K. Haematological, pathological and certain biochemical changes in experimentally infected indigenous with duck plague virus [J]. Indian Journal of Animal Sciences,1983,53:145-149.
[212]Proctor, S.J. Pathogenesis of digestive tract lesions in duck plague[J]. Vet Pathol,1975,12 (5-6) 349-361.
[213]kaszanyitzky, E.J.,Tanyi, J. Studies on the laboratory diagnosis and epizootiology of duck virus hepatitis[J]. Magy Allatorv Lapja,1980,35:808-814.
[214]Friend, M.,Trainer, D.O. Experimental duck virus hepatitis in the mallard[J]. Avian Dis,1972,16 (4):692-699.
[215]Sturm-Ramirez, K.M.,Ellis, T.,Bousfield, B.et al. Reemerging H5N1 influenza viruses in hong kong in 2002 are highly pathogenic to ducks[J]. J Virol.2004,78 (9):4892-4901.
[216]Cooley, A.J.,Van Campen, H.,Philpott, M.S.et al. Pathological lesions in the lungs of ducks infected with influenza a viruses[J]. Vet Pathol,1989,26 (1):1-5.
[217]Zapata, A.G.,Cooper, E.L. The immune system Comparative histophysiology[M]. Micheser:John Wiley and Sona,1991.
[218]Duncan, R. The low pH-dependent entry of avian reovirus is accompanied by two specific cleavages of the major outer capsid protein & mu;2c[J]. Virology,1996,219 (1):179-189.
[219]Labrada, L.,Bodelon, G.,Vinuela, J.et al. Avian reoviruses cause apoptosis in cultured cells: Viral uncoating, but not viral gene expression, is required for apoptosis induction[J]. J Virol,2002,76 (16):7932-7941.
[220]殷震,刘景华.动物病毒学[M].第二版.北京：科学出版社,1997.
[221]赵亚力,孙衍庆,周捷.外源3型呼肠孤病毒污染原代地鼠肾细胞的光景及电镜诊断[J].中国生物制品学杂志,2001,14(1)：33-35.
[222]Touris-Otero, F.,Cortez-San Martin, M.,Martinez-Costas, J.et al. Avian reovirus morphogenesis occurs within viral factories and begins with the selective recruitment of sigmans and lambdaa to microns inclusions[J]. J Mol Biol,2004,341 (2):361-374.
[223]Touris-Otero, F.,Martinez-Costas, J.,Vakharia, V.N.et al. Avian reovirus nonstructural protein microns forms viroplasm-like inclusions and recruits protein sigmans to these structures [J]. Viro]ogy,2004,319 (1):94-106.
[224]Xu, W.,Patrick, M.K.,Hazelton, P.R.et al. Avian reovirus temperature-sensitive mutant tsal2 has a lesion in major core protein sigmaa and is defective in assembly [J]. J Virol,2004,78 (20):11142-11151.
[225]黄灿华,张建红,高玮.应用光景和电镜对病虾组织细胞病理变化的观察与分析[J].中国病毒学,1997,12(4)：364-370.
[226]Alberts, E. Endoplasmic reticulum:Structure and function molecular biology of the cell[M]. Garland:Garland publishing,1994.
[227]Green, D.E. Mitochondria-structure, function, and replication[J]. N Engl J Med,1983,309 (3):182-183.
[228]Griffiths, E.J. Mitochondria-potential role in cell life and death[J]. Cardiovasc Res,2000,46 (1):24-27.
[229]Bielefeldt Ohmann, H.,Bloch, B. Electron microscopic studies of bovine viral diarrhea virus in tissues of diseased calves and in cell cultures[J]. Arch Virol,1982,71 (1):57-74.
[230]Duncan, R.,Chung, K.L.,Faulkner, P. Analysis of a mutant of autographa California nuclear polyhedrosis virus with a defect in the morphogenesis of the occlusion body macromolecular lattice[J]. J Gen Virol,1983,64 (Pt 7) (1531-1542.
[231]Young, L.S.,Dawson, C.W.,Eliopoulos, A.G. Viruses and apoptosis[J]. Br Med Bull,1997,53 (3):509-521.
[232]Koyama, A.H.,Irie, H.,Fukumori, T.et al. Role of virus-induced apoptosis in a host defense mechanism against virus infection[J]. J Med Invest,1998,45 (1-4):37-45.
[233]Teodoro, J.G.,Branton, P.E. Regulation of apoptosis by viral gene products[J]. J Virol,1997,71 (3):1739-1746.
[234]Clarke, P.,Tyler, K.L. Reovirus-induced apoptosis:A minireview[J]. Apoptosis,2003,8 (2):141-150.
[235]Clarke, P.,Debiasi, R.L.,Goody, R.et al. Mechanisms of reovirus-induced cell death and tissue injury:Role of apoptosis and virus-induced perturbation of host-cell signaling and transcription factor activation[J]. Viral Immunol,2005,18 (1):89-115.
[236]Chulu, J.L.,Lee, L.H.,Lee, Y.C.et al. Apoptosis induction by avian reovirus through p53 and mitochondria-mediated pathway[J]. Biochem Biophys Res Commun,2007,356 (3):529-535.
[237]Lin, H.Y.,Chuang, S.T.,Chen, Y.T.et al. Avian reovirus-induced apoptosis related to tissue injury[J].Avian Pathol,2007,36 (2):155-159.
[238]Kerr, J.F.,Wyllie, A.H.,Currie, A.R. Apoptosis:A basic biological phenomenon with wide-ranging implications in tissue kinetics[J]. Br J Cancer,1972,26 (4):239-257.
[239]Fadeel, B.,Orrenius, S. Apoptosis:A basic biological phenomenon with wide-ranging implications in human disease[J]. J Intern Med,2005,258 (6):479-517.
[240]Oppenheim, R.W. Cell death during development of the nervous system[J]. Annu Rev Neurosci,1991,14:453-501.
[241]Koyama, A.H.,Arakawa, T.,Adachi, A. Characterization of apoptosis induced by sorbitol:A unique system for the detection of antiapoptotic activities of viruses[J]. Microbes Infect,2000,2 (6):599-606.
[242]Miller, L.C.,Fox, J.M. Apoptosis and porcine reproductive and respiratory syndrome virus[J]. Vet Immunol Immunopathol,2004,102 (3):131-142.
[243]Wyllie, A.H.,Kerr, J.F.,Currie, A.R. Cell death:The significance of apoptosis[J]. Int Rev Cytol,1980,68:251-306.
[244]Kerr J.F.R., W.C.M., Harmon B.V.. Apoptosis. Its significance in cancer and cancer therapy [J]. Cancer,1994,73:2013-2026.
[245]Oberhammer F.A., H.K., Fr6sehl G, et al.. Chromatin condensation during apoptosis is accompanied by degradation of lamin a+b, without enhanced activation of cdc2 kinasefJ]. The Journal of Cell Biology,1994,126 (4):827-837.
[246]Strasser, A.,O'Connor, L.,Dixit, V.M. Apoptosis signaling[J]. Annu Rev Biochem,2000,69: 217-245.
[247]Mangili, F.,Cigala, C.,Santambrogio, G. Staining apoptosis in paraffin sections. Advantages and limits[J]. Anal Quant Cytol Histol,1999,21 (3):273-276.
[248]Saraste, A. Morphologic criteria and detection of apoptosis[J]. Herz,1999,24 (3):189-195.
[249]Rihova, Z.,Sefc, L.,Necvas, E. [methods used in the detection of apoptosis][J]. Cas Lek Cesk,2001,140 (6):168-172.
[250]Vaux, D.L.,Haecker, G.,Strasser, A. An evolutionary perspective on apoptosis[J]. Cell,1994,76 (5):777-779.
[251]Jaworowski, A.,Crowe, S.M. Does hiv cause depletion of CD4+t cells in vivo by the induction of apoptosis?[J]. Immunol Cell Biol,1999,77 (1):90-98.
[252]Ito, T.,Kobayashi, Y.,Morita, T.et al. Virulent influenza a viruses induce apoptosis in chickens[J]. Virus Res,2002,84 (1-2):27-35.
[253]Jeurissen, S.H.,Wagenaar, F.,Pol, J.M.et al. Chicken anemia virus causes apoptosis of thymocytes after in vivo infection and of cell lines after in vitro infection[J]. J Virol,1992,66 (12):7383-7388.
[254]Rosenberger, J.K.,Sterner, F.J.,Botts, S.et al. In vitro and in vivo characterization of avian reoviruses. I. Pathogenicity and antigenic relatedness of several avian reovirus isolates[J]. Avian Dis,1989,33 (3):535-544.
[255]DeBiasi, R.L.,Robinson, B.A.,Sherry, B.et al. Caspase inhibition protects against reovirus-induced myocardial injury in vitro and in vivo[J]. J Virol,2004,78 (20):11040-11050.
[256]Salako, M.A.,Carter, M.J.,Kass, G.E. Coxsackievirus protein 2bc blocks host cell apoptosis by inhibiting caspase-3[J]. J Biol Chem,2006,281 (24):16296-16304.
[257]Everett, H.,McFadden, G. Apoptosis:An innate immune response to virus infection[J]. Trends Microbiol,1999,7 (4):160-165.
[258]Dockrell, D.H.,Badley, A.D.,Villacian, J.S.et al. The expression of fas ligand by macrophages and its upregulation by human immunodeficiency virus infection[J]. J Clin Invest,1998,101 (11):2394-2405.
[259]Sabri, F.,Chiodi, F.,Piret, J.P.et al. Soluble factors released by virus specific activated cytotoxic t-lymphocytes induce apoptotic death of astroglioma cell lines[J]. Brain Pathol,2003,13 (2):165-175.
[260]Kawanishi, M. Epstein-barr virus bhrfl protein protects intestine 407 epithelial cells from apoptosis induced by tumor necrosis factor alpha and anti-fas antibody[J]. J Virol,1997,71 (4):3319-3322.
[261]Hay, S.,Kannourakis, G. A time to kill:Viral manipulation of the cell death program[J]. J Gen Virol,2002,83 (Pt7):1547-1564.
[262]Hetts, S.W. To die or not to die:An overview of apoptosis and its role in disease[J]. JAMA,1998,279 (4):300-307.
[263]Shih, W.L.,Hsu, H.W.,Liao, M.H.et al. Avian reovirus sigmac protein induces apoptosis in cultured cells[J].Virology,2004,321 (1):65-74.
[264]陈志胜,马春全,卢玉葵,等.番鸭呼肠孤病毒诱导雏番鸭免疫器官细胞凋亡的研究[J].中国预防兽医学报,2005,27(6)：457-458.
[265]Zamzami, N.,Marchetti, P.,Castedo, M.et al. Reduction in mitochondrial potential constitutes an early irreversible step of programmed lymphocyte death in vivo[J]. J Exp Med,1995,181 (5):1661-1672.
[266]McConkey, D.J.,Nicotera, P.,Orrenius, S. Signalling and chromatin fragmentation in thymocyte apoptosis[J]. Immunol Rev,1994,142:343-363.
[267]Newmeyer, D.D.,Farschon, D.M.,Reed, J.C. Cell-free apoptosis in xenopus egg extracts: Inhibition by Bcl-2 and requirement for an organelle fraction enriched in mitochondria[J]. Cell,1994,79 (2):353-364.
[268]Lee, C.W. Pathogenesis of infectious bronchitis virus with different routes of inoculation and the effect of in vivo serial passage in nephropathogenicity using cloacal infection[J]. Korean Journal of Veterinary Service,2002,25 (1):87-96.
[269]Psalla, D.,Psychas, V.,Spyrou, Vet al. Pathogenesis of experimental encephalomyocarditis: A histopathological, immunohistochemical and virological study in mice[J]. J Comp Pathol,2006,135 (2-3):142-145.
[270]许益民.免疫过氧化物酶染色方法[M].北京：中国农业出版社,2001.
[271]汪家政,范明.蛋白质技术手册[M].北京：科学出版社,2000.
[272]Temponi M., K.T., Perosa F., et al. Purification of murine igg monoclonal antibodies by precipitation with caprylic acid:Comparison with other methods of purification[J]. Hybridoma,1989,8 (1):85-95.
[273]McGuire J.M., D.M., Smith K.D.. The resolution of the neutral n-linked oligosaccharides of igg by high ph anion-exchange chromatography[J]. Carbohydr. Res.,1996,292 (1-9.
[274]John, E.C.精编蛋白质科学实验指南[M].北京：科学出版社,2007.
[275]Sambrook, J., Russell, D. W. Molecular cloning:A. Laboratory manual. [M]. New York:Cold Spring Harbor:Cold Spring Harbor Laboratory Press,2001.
[276]Harlow E.抗体技术实验指南[M].北京：科学出版社,2002.
[277]蔡文琴,王伯法.实用免疫细胞化学与核酸分子杂交技术[M].成都：四川科学技术出版社,1994.
[278]陈立功,刘向平,武现军,等.免疫组化法检测鹅源新城疫病毒在人工感染鹅体内的动态分布[J].中国兽医杂志,2009,(9)：15-17.
[279]Ramos-Vara, J.A.,Beissenherz, M.E. Optimization of immunohistochemical methods using two different antigen retrieval methods on formalin-fixed paraffin-embedded tissues:Experience with 63 markers[J]. J Vet Diagn Invest,2000,12 (4):307-311.
[280]Shi, S.R.,Key, M.E.,Kalra, K.L. Antigen retrieval in formalin-fixed, paraffin-embedded tissues:An enhancement method for immunohistochemical staining based on microwave oven heating of tissue sections[J]. J Histochem Cytochem,1991,39 (6):741-748.
[281]Kahveci, Z.,Minbay, F.Z.,Noyan, S.et al. A comparison of microwave heating and proteolytic pretreatment antigen retrieval techniques in formalin fixed, paraffin embedded tissues[J]. Biotech Histochem,2003,78 (2):119-128.
[282]Hoetelmans, R.W.,van Slooten, HJ.,Keijzer, R.et al. Comparison of the effects of microwave heating and high pressure cooking for antigen retrieval of human and rat be 1-2 protein in formaldehyde-fixed, paraffin-embedded sections[J]. Biotech Histochem,2002,77 (3):137-144.
[283]Gao, C.,Wang, A.Y.,Han, Y.J. Microwave antigen retrieval blocks endogenous peroxidase activity in immunohistochemistry[J]. Appl Immunohistochem Mol Morphol,2008,16 (4):393-399.
[284]Evke, E.,Minbay, F.Z.,Temel, S.Get al. Immunohistochemical detection of p53 protein in basal cell skin cancer after microwave-assisted antigen retrieval [J]. J Mol Histol,2009,40 (1):13-21.
[285]Shi, S.R.,Cote, R.J.,Taylor, C.R. Antigen retrieval immunohistochemistry:Past, present, and future[J].J Histochem Cytochem,1997,45 (3):327-343.
[286]Ramos-Vara, J.A. Technical aspects of immunohistochemistry [J]. Vet Pathol,2005,42 (4):405-426.
[287]Shawky, S. Target cells for duck enteritis virus in lymphoid organs[J]. Avian Pathol,2000,29(6)：609-616.
[288]程安春,韩晓英,汪铭书,等.用间接免疫荧光染色法检测鸭病毒性肠炎病毒在人工感染鸭体内的侵染过程和分布规律[J].畜牧兽医学报,2007,38(9)：942-946.
[289]程国富,胡薛英,周诗其,等.单克隆抗体PAP法对实验感染雏鸭体内鸭肝炎病毒的定位检测[J].华中农业大学学报,1996,15(6)：568-572.
[290]汪铭书,程安春,陈孝跃.鸭病毒性肝炎研究一强毒在雏鸭和成年鸭体内的分布和排泄[J].中国兽医学报,1997,17(3)：254-257.
[291]郑林英.间接夹心ELISA检测鸭肿头败血症病毒的研究[D].雅安：四川农业大学2005.
[292]陈元坤.鸡病毒性关节炎病毒增殖特性与间接ELISA抗体检测方法的建立研究[D].雅安：四川农业大学,2007.
[293]朱立平,陈学清.免疫学常用实验方法[M].北京：人民医学出版社,2000.
[294]McLaren, M.L.,Lillywhite, J.E.,Au, A.C. Indirect enzyme linked immunosorbent assay (ELISA):Practical aspects of standardization and quality control[J]. Med Lab Sci,1981,38 (3):245-251.
[295]Desquesnes, M. [international and regional standardization of immunoenzyme tests: Methods, concerns and limitations][J]. Rev Sci Tech,1997,16 (3):809-823.
[296]Hansson, A.S.,Heinegard, D.,Piette, J.C.et al. The occurrence of autoantibodies to matrilin 1 reflects a tissue-specific response to cartilage of the respiratory tract in patients with relapsing polychondritis[J]. Arthritis Rheum,2001,44 (10):2402-2412.
[297]Lim, K.J.,Park, J.W.,Sohn, M.J.et al. A direct sandwich elisa to detect antibodies against the c-terminal region of merozoite surface protein 1 could be a useful diagnostic method to identify plasmodium vivax exposed persons[J]. Parasitol Res,2002,88 (9):855-860.
[298]Kishimoto, T.,Ando, S.,Numazaki, K.et al. Assay of chlamydia pneumoniae-specific igm antibodies by elisa method-reduction of non-specific reaction and resetting of serological criteria by measuring igm antibodies[J]. Jpn J Infect Dis,2009,62 (4):260-264.