犬细小病毒的分离鉴定及动物感染模型的建立
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摘要
犬细小病毒病是一种具有高度接触性的烈性传染病，犬细小病毒病以呕吐、腹泻和白细胞减少为特征。自然感染犬细小病毒的病例经常发生，即使CPV疫苗免疫后也会发病，幼犬的死亡率非常高，随着犬饲养业的快速发展，特别是实验用犬和宠物犬饲养量的大幅增加，犬感染细小病毒病日趋严重，给犬饲养业造成重大的经济损失。犬细小病毒（CPV）是一条单股负链线性的DNA病毒，无囊膜，对物理化学因素具有很强的抵抗力，病毒在粪便中可以存活很长时间，致病力不会降低，病毒大量存在于粪便中，传染性也最强。病犬是主要的传染源，动物主要是通过接触感染发病。
本实验收集长春地区患有严重传染性腹泻病死犬的临床样本，通过犬细小病毒核酸PCR鉴定后,筛选阳性样本,用F81细胞进行犬细小病毒的分离，结果从5份核酸PCR鉴定犬细小病毒阳性样本中分离得到4株病毒，并通过电镜观察、血凝试验、细胞感染谱鉴定后确定所分离的毒株为犬细小病毒株，并分别命名为：CPV-BM、CPV-LTS、CPV-GYW、CPV-GZL。
为确定长春地区犬细小流行株的基因型，以更好地进行预防和治疗，对本实验所分离的毒株和实验室保存的毒株进行病毒VP2基因的PCR扩增和序列测定，通过序列分析显示，本实验分离的细小病毒株CPV-BM、CPV-LTS、CPV-GYW、CPV-GZL的VP2蛋白第426位氨基酸是Asn，为细小病毒CPV-2a型，同时与实验室早期分离并保存毒株CPV-DM比较，可以推断细小病毒正不断发生进化。
本研究从病毒感染性出发，以不同感染方式为切入点，感染实验动物，建立犬细小病毒病的感染模型。本研究对抗病毒药物的研发提供必要的动物模型，为评价抗病毒新药物疗效做准备。本实验使用细小病毒分离株CPV-BM，分别通过口服、滴鼻点眼、皮下注射、肌肉注射等四种不同途径感染2-3月龄普通犬。感染后，每天观察记录犬的临床症状，并检测排毒状况，并检测感染后第2、4、6、8、10、12天血液白细胞数值变化。结果显示：口服途径感染后，实验犬最先表现出临床症状，在感染第2天平均体温开始升高，第5天平均体重出现下降，3例出现便血，发病率100%，死亡率达75%，病程为6～8天，白细胞数值最低为2.32×109/L；滴鼻点眼途径感染后，实验犬也表现出精神沉郁，脱水等临床症状，但症状出现时间较口服途径感染组晚，在感染后第4天平均体温升高，同时于第6天发病犬体重开始下降，1例出现便血，发病率75%，死亡率25%，白细胞数值最低为4.8×109/L；皮下注射和肌肉注射途径感染后，均未表现出明显的临床症状。通过实验证明，所建立的细小病毒感染模型以口服途径感染效果最好，具有典型的临床症状，且发病率100%。为犬细小病毒疫苗的研制和治疗药物的评价提供了稳定可靠的动物模型。
The canine parvovirus is a deadly infection disease which characterized by acutevomiting, diarrhea and leukopenia. Natural cases of canine parvirus infection oftenoccurs.Though the CPV vaccine was used in disease, the incidence of puppy mortalityis highly. In recent years, with a substantial increase in raising the amount ofexperiments dogs and pet dogs, the Canine parvovirus are becoming serious, it bringsignificantly losses to the breeding industry. CPV is a single-stranded negative linearDNA virus, has a strong resistance to physical and chemical factors. The dog is themainly source of infection, especially in contact with the feces of dogs, because thevirus could survive in feces for a long time, and the pathogenicity isn’t reduce. Thevirus has a high level in feces, and the infectious is also the strongest.
We collect the clinical samples from infectious diarrhea dogs in Chang-chun, weidentified the samples by PCR, and separated the virus by F81cell. The result showswe separated four CPV strains from five CPV positive samples through microscopy,hemagglutination and cell infected with spectrum test. At last, we named the fourstrains as CPV-BM, CPV-LTS, CPV-GYW, CPV-GZL.
To determine the gene-type of the CPV trend in Changchun, prevent and treat theanimals better, we amplicated and determined the sequence of CPV VP2gene. Thesequence analysis showed that the426amino acids of the four strains is Asn, theybelong to CPV-2a. The mainly type of CPV is CPV-2a in Changchun. CPV-DM is anearly strains, compared to CPV-BM, CPV-LTS, CPV-GYW, CPV-GZL is lowhomology, and amino acid differences are larger. In recent years, CPV strains are stillevolving.
My study started from the infection of virus and different patterns of infection,isolated the virus from the diagnosis of dogs, identified and analysised it. We infectedin experimental animals and established a model of canine parvirus infection. Thestudy provide the necessary animal models for the development of antivirus drugs,and prepare for the evaluation of new antiviral drug dfficacy. To provide the animal model for CPV vaccine development and treatment of drug evaluation, Theexperimental dogs are ordinary pet dogs which is2-3months old and the CPV test isnegative, select BM strains as the attack-virus, Performing clinical, hematological,serological, virus isolation and pathological by four methods including oralapplication, respiration and eye-drop application, intramuscular injection andhypodermic injection, the dose is1ml/kg, we collected fecal to detect the discharge ofthe virus, tested the blood changes on2,4,6,8,10,12days.
The results shows the oral group is the first to occur the clinical symptoms whichshows depression, anorexia, weight loss, dehydration and so on. The averagetemperature increased from the next day to the second day, the average weightdeclined on the fifth day, three dogs appeared hemorrhagic in the fecal,100%of thewhole group is sick, and the mortality rate is75%. The course of the disease is6-8d.The lowest WBC was2.32×109/L; The eyedrop application group also shows theclinical symptoms after infection virus, but the time is late than the oral group, Theaverage temperature increased from the fourth day to the sixth day, the average weightdeclined on the sixth day, one dog appeared hemorrhagic in the fecal, the disease rateis75%, the the mortality rate is25%. The lowest WBC was4.8×109/L. Theintramuscular injectionand hypodermic injection group didn’t show clinical symptoms.The result shows that oral infection is the best pathway in the establishment of theinfection, with typical clinical symptoms, and the incidence is100%. Our studyprovide a stable animal model for the evaluation of CPV vaccines and treatmentdrugs.
本实验收集长春地区患有严重传染性腹泻病死犬的临床样本，通过犬细小病毒核酸PCR鉴定后,筛选阳性样本,用F81细胞进行犬细小病毒的分离，结果从5份核酸PCR鉴定犬细小病毒阳性样本中分离得到4株病毒，并通过电镜观察、血凝试验、细胞感染谱鉴定后确定所分离的毒株为犬细小病毒株，并分别命名为：CPV-BM、CPV-LTS、CPV-GYW、CPV-GZL。
为确定长春地区犬细小流行株的基因型，以更好地进行预防和治疗，对本实验所分离的毒株和实验室保存的毒株进行病毒VP2基因的PCR扩增和序列测定，通过序列分析显示，本实验分离的细小病毒株CPV-BM、CPV-LTS、CPV-GYW、CPV-GZL的VP2蛋白第426位氨基酸是Asn，为细小病毒CPV-2a型，同时与实验室早期分离并保存毒株CPV-DM比较，可以推断细小病毒正不断发生进化。
本研究从病毒感染性出发，以不同感染方式为切入点，感染实验动物，建立犬细小病毒病的感染模型。本研究对抗病毒药物的研发提供必要的动物模型，为评价抗病毒新药物疗效做准备。本实验使用细小病毒分离株CPV-BM，分别通过口服、滴鼻点眼、皮下注射、肌肉注射等四种不同途径感染2-3月龄普通犬。感染后，每天观察记录犬的临床症状，并检测排毒状况，并检测感染后第2、4、6、8、10、12天血液白细胞数值变化。结果显示：口服途径感染后，实验犬最先表现出临床症状，在感染第2天平均体温开始升高，第5天平均体重出现下降，3例出现便血，发病率100%，死亡率达75%，病程为6～8天，白细胞数值最低为2.32×109/L；滴鼻点眼途径感染后，实验犬也表现出精神沉郁，脱水等临床症状，但症状出现时间较口服途径感染组晚，在感染后第4天平均体温升高，同时于第6天发病犬体重开始下降，1例出现便血，发病率75%，死亡率25%，白细胞数值最低为4.8×109/L；皮下注射和肌肉注射途径感染后，均未表现出明显的临床症状。通过实验证明，所建立的细小病毒感染模型以口服途径感染效果最好，具有典型的临床症状，且发病率100%。为犬细小病毒疫苗的研制和治疗药物的评价提供了稳定可靠的动物模型。
The canine parvovirus is a deadly infection disease which characterized by acutevomiting, diarrhea and leukopenia. Natural cases of canine parvirus infection oftenoccurs.Though the CPV vaccine was used in disease, the incidence of puppy mortalityis highly. In recent years, with a substantial increase in raising the amount ofexperiments dogs and pet dogs, the Canine parvovirus are becoming serious, it bringsignificantly losses to the breeding industry. CPV is a single-stranded negative linearDNA virus, has a strong resistance to physical and chemical factors. The dog is themainly source of infection, especially in contact with the feces of dogs, because thevirus could survive in feces for a long time, and the pathogenicity isn’t reduce. Thevirus has a high level in feces, and the infectious is also the strongest.
We collect the clinical samples from infectious diarrhea dogs in Chang-chun, weidentified the samples by PCR, and separated the virus by F81cell. The result showswe separated four CPV strains from five CPV positive samples through microscopy,hemagglutination and cell infected with spectrum test. At last, we named the fourstrains as CPV-BM, CPV-LTS, CPV-GYW, CPV-GZL.
To determine the gene-type of the CPV trend in Changchun, prevent and treat theanimals better, we amplicated and determined the sequence of CPV VP2gene. Thesequence analysis showed that the426amino acids of the four strains is Asn, theybelong to CPV-2a. The mainly type of CPV is CPV-2a in Changchun. CPV-DM is anearly strains, compared to CPV-BM, CPV-LTS, CPV-GYW, CPV-GZL is lowhomology, and amino acid differences are larger. In recent years, CPV strains are stillevolving.
My study started from the infection of virus and different patterns of infection,isolated the virus from the diagnosis of dogs, identified and analysised it. We infectedin experimental animals and established a model of canine parvirus infection. Thestudy provide the necessary animal models for the development of antivirus drugs,and prepare for the evaluation of new antiviral drug dfficacy. To provide the animal model for CPV vaccine development and treatment of drug evaluation, Theexperimental dogs are ordinary pet dogs which is2-3months old and the CPV test isnegative, select BM strains as the attack-virus, Performing clinical, hematological,serological, virus isolation and pathological by four methods including oralapplication, respiration and eye-drop application, intramuscular injection andhypodermic injection, the dose is1ml/kg, we collected fecal to detect the discharge ofthe virus, tested the blood changes on2,4,6,8,10,12days.
The results shows the oral group is the first to occur the clinical symptoms whichshows depression, anorexia, weight loss, dehydration and so on. The averagetemperature increased from the next day to the second day, the average weightdeclined on the fifth day, three dogs appeared hemorrhagic in the fecal,100%of thewhole group is sick, and the mortality rate is75%. The course of the disease is6-8d.The lowest WBC was2.32×109/L; The eyedrop application group also shows theclinical symptoms after infection virus, but the time is late than the oral group, Theaverage temperature increased from the fourth day to the sixth day, the average weightdeclined on the sixth day, one dog appeared hemorrhagic in the fecal, the disease rateis75%, the the mortality rate is25%. The lowest WBC was4.8×109/L. Theintramuscular injectionand hypodermic injection group didn’t show clinical symptoms.The result shows that oral infection is the best pathway in the establishment of theinfection, with typical clinical symptoms, and the incidence is100%. Our studyprovide a stable animal model for the evaluation of CPV vaccines and treatmentdrugs.
引文
[1]李河林,焦铁军,刘淑红等.犬细小病毒病研究进展[J].动物医学进展,2008,05):73-7.
[2]周云朵.犬细小病毒的分离鉴定与生物学特性研究[D];华中农业大学,2011.
[3] CHINCHKAR S R, MOHANA SUBRAMANIAN B, HANUMANTHA RAO N,et al. Analysis of VP2gene sequences of canine parvovirus isolates in India [J]. ArchVirol,2006,151(9):1881-7.
[4] DOKI M, FUJITA K, MIURA R, et al. Sequence analysis of VP2gene of canineparvovirus isolated from domestic dogs in Japan in1999and2000[J]. CompImmunol Microbiol Infect Dis,2006,29(4):199-206.
[5] MCKENNA R, OLSON N H, CHIPMAN P R, et al. Three-dimensional structureof Aleutian mink disease parvovirus: implications for disease pathogenicity [J]. JVirol,1999,73(8):6882-91.
[6] FILIPOV C, DECARO N, DESARIO C, et al. Canine parvovirus epidemiologyin Bulgaria [J]. J Vet Diagn Invest,2011,23(1):152-4.
[7] ZHAO J, YAN X, WU W.[Origin and evolution of canine parvovirus--a review][J]. Wei Sheng Wu Xue Bao,2011,51(7):869-75.
[8] MOCHIZUKI M, OHSHIMA T, UNE Y, et al. Recombination between vaccineand field strains of canine parvovirus is revealed by isolation of virus in canine andfeline cell cultures [J]. J Vet Med Sci,2008,70(12):1305-14.
[9] LARSON L J, SCHULTZ R D. Do two current canine parvovirus type2and2bvaccines provide protection against the new type2c variant?[J]. Vet Ther,2008,9(2):94-101.
[10]JEOUNG S Y, AHN S J, KIM D.Genetic analysis of VP2gene of canineparvovirus isolates in Korea [J]. J Vet Med Sci,2008,70(7):719-22.
[11] DECARO N,DESARIO C,ADDIE D D, et al. The study molecular epidemiologyof canine parvovirus, Europe [J]. Emerg Infect Dis,2007,13(8):1222-4.
[12]DECARO N, DESARIO C, PARISI A,et al. Genetic analysis of canine parvovirustype2c [J]. Virology,2009,385(1):5-10.
[13]KANG B K, SONG D S, LEE C S, et al. Prevalence and genetic characterizationof canine parvoviruses in Korea [J]. Virus Genes,2008,36(1):127-33.
[14]DECARO N, DESARIO C, ELIA G, et al. Evidence for immunisation failure invaccinated adult dogs infected with canine parvovirus type2c [J]. New Microbiol,2008,31(1):125-30.
[15]CAVALLI A, MARTELLA V, DESARIO C, et al. Evaluation of the antigenicrelationships among canine parvovirus type2variants [J]. Clin Vaccine Immunol,2008,15(3):534-9.
[16]JOAO VIEIRA M, SILVA E, OLIVEIRA J, et al. Canine parvovirus2c infectionin central Portugal [J]. J Vet Diagn Invest,2008,20(4):488-91.
[17]SIEDEK E M, SCHMIDT H, STURE G H, et al. Vaccination with canineparvovirus type2(CPV-2) protects against challenge with virulent CPV-2b andCPV-2c [J]. Berl Munch Tierarztl Wochenschr,2011,124(1-2):58-64.
[18]TOUIHRI L, BOUZID I, DAOUD R, et al. Molecular characterization of canineparvovirus-2variants circulating in Tunisia [J]. Virus Genes,2009,38(2):249-58.
[19]HARBISON C E, LYI S M, WEICHERT W S, et al. Early steps in cell infectionby parvoviruses: host-specific differences in cell receptor binding but similarendosomal trafficking [J]. J Virol,2009,83(20):10504-14.
[20]OHSHIMA T, MOCHIZUKI M. Evidence for recombination between felinepanleukopenia virus and canine parvovirus type2[J]. J Vet Med Sci,2009,71(4):403-8.
[21]VIHINEN-RANTA M, KALELA A, MAKINEN P, et al. Intracellular route ofcanine parvovirus entry [J]. J Virol,1998,72(1):802-6.
[22]PARKER J S, PARRISH C R. Canine parvovirus host range is determined by thespecific conformation of an additional region of the capsid [J]. J Virol,1997,71(12):9214-22.
[23]HOELZER K, SHACKELTON L A, PARRISH C R,et al. Phylogenetic analysisreveals the emergence, evolution and dispersal of carnivore parvoviruses [J]. J GenVirol,2008,89(Pt9):2280-9.
[24]CALDERON M G,ROMANUTTI C, A D A,et al. Evolution of canine parvovirusin Argentina between years2003and2010: CPV2c has become the predominantvariant affecting the domestic dog population [J]. Virus Res,2011,157(1):106-10.
[25]贺英,曹旺斌,史秋梅等.基于转铁蛋白受体对犬细小病毒原代细胞组织嗜性的研究,中国黑龙江齐齐哈尔, F,2010[C].
[26]PARKER J S, PARRISH C R. Cellular uptake and infection by canine parvovirusinvolves rapid dynamin-regulated clathrin-mediated endocytosis, followed by slowerintracellular trafficking [J]. J Virol,2000,74(4):1919-30.
[27]耿志贤,时彦胜,郑亚平等.犬细小病毒病的流行现状调查[J].中国畜牧兽医,2009,11):135-7.
[28]BATTILANI M, BALBONI A, USTULIN M, et al. Genetic complexity andmultiple infections with more Parvovirus species in naturally infected cats [J]. Vet Res,2011,42(1):43.
[29]VIHINEN-RANTA M, YUAN W, PARRISH C R. Cytoplasmic trafficking of thecanine parvovirus capsid and its role in infection and nuclear transport [J]. J Virol,2000,74(10):4853-9.
[30]NTAFIS V, XYLOURI E, KALLI I, et al. Characterization of Canine parvovirus2variants circulating in Greece [J]. J Vet Diagn Invest,2010,22(5):737-40.
[31]MEERS J, KYAW-TANNER M, BENSINK Z, et al. Genetic analysis of canineparvovirus from dogs in Australia [J]. Aust Vet J,2007,85(10):392-6.
[32]WU Z, SUN H, ZHANG X.[Immunoadjuvant effect of the extracellular domainof canine CTLA-4on the VP2protein of canine parvovirus][J]. Wei Sheng Wu XueBao,2008,48(3):369-74.
[33]HAFENSTEIN S, BOWMAN V D, SUN T, et al. Structural comparison ofdifferent antibodies interacting with parvovirus capsids [J]. J Virol,2009,83(11):5556-66.
[34]CASTRO T X, COSTA E M, LEITE J P, et al. Monitoring of canine parvovirus(CPV) strains detected in vaccinated puppies in Brazil [J]. Res Vet Sci,2011,90(2):336-40.
[35]TAGUCHI M, NAMIKAWA K, MARUO T, et al. Antibody titers for canineparvovirus type-2, canine distemper virus, and canine adenovirus type-1in adulthousehold dogs [J]. Can Vet J,2011,52(9):983-6.
[36]贺英,张曼夫,曹旺斌.犬细小病毒宿主范围和入侵途径研究进展[J].畜牧与兽医,2009,01):101-4.
[37]SPIBEY N, GREENWOOD N M, SUTTON D, et al. Canine parvovirus type2vaccine protects against virulent challenge with type2c virus [J]. Vet Microbiol,2008,128(1-2):48-55.
[38]MARULAPPA S Y, KAPIL S. Simple tests for rapid detection of canineparvovirus antigen and canine parvovirus-specific antibodies [J]. Clin VaccineImmunol,2009,16(1):127-31.
[39]ELIA G, CAVALLI A, DESARIO C, et al. Detection of infectious canineparvovirus type2by mRNA real-time RT-PCR [J]. J Virol Methods,2007,146(1-2):202-8.
[40]宋永奇,李玉燕,吕艳丽等.3种犬细小病毒感染诊断方法比较[J].畜牧与兽医,2008,06):31-5.
[41]郭伟.犬细小病毒分离鉴定及环介导等温扩增快速检测方法的建立[D];内蒙古农业大学,2010.
[42]GODSALL S A, CLEGG S R, STAVISKY J H, et al. Epidemiology of canineparvovirus and coronavirus in dogs presented with severe diarrhoea to PDSA PetAidhospitals [J]. Vet Rec,2010,167(6):196-201.
[43]CHO H S, KANG J I, PARK N Y. Detection of canine parvovirus in fecalsamples using loop-mediated isothermal amplification [J]. J Vet Diagn Invest,2006,18(1):81-4.
[44]DECARO N, MARTELLA V, DESARIO C, et al. First detection of canineparvovirus type2c in pups with haemorrhagic enteritis in Spain [J]. J Vet Med BInfect Dis Vet Public Health,2006,53(10):468-72.
[45]杨颗粒,张丰范,孙玉彬.犬细小病毒病的诊断和治疗[J].农村科技,1995,07):21.
[46]CLEGG S R, COYNE K P, PARKER J, et al. Molecular epidemiology andphylogeny reveal complex spatial dynamics in areas where canine parvovirus isendemic [J]. J Virol,2011,85(15):7892-9.
[47]BRAGG R F, DUFFY A L, DECECCO F A, et al. Clinical evaluation of a singledose of immune plasma for treatment of canine parvovirus infection [J]. J Am VetMed Assoc,2012,240(6):700-4.
[48]孙艳芳,贾宁,扎西英派等.犬细小病毒病的病理学观察; proceedings of the中国畜牧兽医学会兽医病理学分会第十五次、中国病理生理学会动物病生专业委员会第十四次学术讨论会,中国黑龙江哈尔滨, F,2007[C].
[49]ZHANG R, YANG S, ZHANG W, et al. Phylogenetic analysis of the VP2gene ofcanine parvoviruses circulating in China [J]. Virus Genes,2010,40(3):397-402.
[50]MAJER-DZIEDZIC B, JAKUBCZAK A, ZIETEK J. Phylogenetic analysis ofcanine parvovirus CPV-2strains and its variants isolated in Poland [J]. Pol J Vet Sci,2011,14(3):379-84.
[51]任宝波,孙冰.犬细小病毒感染防治的研究进展[J].养殖技术顾问,2007,04):60-1.
[52]YOON S H, JEONG W, KIM H J, et al. Molecular insights into the phylogeny ofcanine parvovirus2(CPV-2) with emphasis on Korean isolates: a Bayesian approach[J]. Arch Virol,2009,154(8):1353-60.
[53]陈琛,邬静,张海亮等.犬细小病毒病诊断与防治情况调查[J].湖北农业科学,2010,08):1926-8.
[54]PRATELLI A, CAVALLI A, NORMANNO G, et al. Immunization of pups withmaternally derived antibodies to canine parvovirus (CPV) using a modified-livevariant (CPV-2b)[J]. J Vet Med B Infect Dis Vet Public Health,2000,47(4):273-6.
[55]王洪林.犬细小病毒北京株分离和动物发病模型建立[D];中国农业科学院,2009.
[56]JIANG W, BAKER H J, SWANGO L J, et al. Nucleic acid immunization protectsdogs against challenge with virulent canine parvovirus [J]. Vaccine,1998,16(6):601-7.
[57]MOHAN RAJ J, MUKHOPADHYAY H K, THANISLASS J, et al. Isolation,molecular characterization and phylogenetic analysis of canine parvovirus [J]. InfectGenet Evol,2010,10(8):1237-41.
[58]DECARO N, BUONAVOGLIA C. Canine parvovirus--a review ofepidemiological and diagnostic aspects, with emphasis on type2c [J]. Vet Microbiol,2012,155(1):1-12.
[59]李晋涛,魏静,牟芝蓉等.人源轮状病毒的小型猪腹泻模型建立[J].中国比较医学杂志,2006,09):577.
[60]廖列如,张林,孔庆波等.犬冠状病毒的分离及鉴定[J].安徽农业科学,2007,30):9549-51+98.
[61]LITSTER A, NICHOLS J, VOLPE A. Prevalence of positive antibody test resultsfor canine parvovirus (CPV) and canine distemper virus (CDV) and response tomodified live vaccination against CPV and CDV in dogs entering animal shelters [J].Vet Microbiol,2011,
[62]邓朝晖,许春娣,陈舜年.新生小鼠轮状病毒腹泻模型建立及病理学致病机制研究[J].临床儿科杂志,2008,10):862-5.
[63]LARSON L J, SCHULTZ R D. Three-year serologic immunity against canineparvovirus type2and canine adenovirus type2in dogs vaccinated with a caninecombination vaccine [J]. Vet Ther,2007,8(4):305-10.
[64]MOON H S, LEE S A, LEE S G, et al. Comparison of the pathogenicity in threedifferent Korean canine parvovirus2(CPV-2) isolates [J]. Vet Microbiol,2008,131(1-2):47-56.
[65]阿拉坦布日古德,刘晓松,张喜彦等.犬细小病毒实验感染研究[J].内蒙古兽医,1995,02):1-4.
[66]IRIS K, LEONTIDES L S, MYLONAKIS M E, et al. Factors affecting theoccurrence, duration of hospitalization and final outcome in canine parvovirusinfection [J]. Res Vet Sci,2010,89(2):174-8.
[67]GOODMAN L B, LYI S M, JOHNSON N C, et al. Binding site on the transferrinreceptor for the parvovirus capsid and effects of altered affinity on cell uptake andinfection [J]. J Virol,2010,84(10):4969-78.
[68]NYKKY J, TUUSA J E, KIRJAVAINEN S, et al. Mechanisms of cell death incanine parvovirus-infected cells provide intuitive insights to developing nanotools formedicine [J]. Int J Nanomedicine,2010,5(417-28.
[69]MUZ D, OGUZOGLU T C, TIMURKAN M O, et al. Characterization of thepartial VP2gene region of canine parvoviruses in domestic cats from Turkey [J].Virus Genes,2012,44(2):301-8.
[70]GALLO CALDERON M, WILDA M, BOADO L, et al. Study of canineparvovirus evolution: comparative analysis of full-length VP2gene sequences fromArgentina and international field strains [J]. Virus Genes,2012,44(1):32-9.
[71]LLAMAS-SAIZ A L, AGBANDJE-MCKENNA M, PARKER J S, et al.Structural analysis of a mutation in canine parvovirus which controls antigenicity andhost range [J]. Virology,1996,225(1):65-71.
[72]CHANG S F, SGRO J Y, PARRISH C R. Multiple amino acids in the capsidstructure of canine parvovirus coordinately determine the canine host range andspecific antigenic and hemagglutination properties [J]. J Virol,1992,66(12):6858-67.
[73]GEMMA T, KAI C, MIKAMI T. Suppression of canine parvovirus growth inCRFK cells by canine distemper virus [J]. J Vet Med Sci,1995,57(3):535-7.
[74]CHAPMAN M S, ROSSMANN M G. Structure, sequence, and functioncorrelations among parvoviruses [J]. Virology,1993,194(2):491-508.
[75]PARRISH C R. Pathogenesis of feline panleukopenia virus and canine parvovirus[J]. Baillieres Clin Haematol,1995,8(1):57-71.
[76]TRUYEN U.[Canine parvovirus: recent knowledge of the origin anddevelopment of a viral pathogen][J]. Tierarztl Prax,1994,22(6):579-84.
[77]TRUYEN U.[Evolution of canine parvovirus: loss and gain of the feline host][J].Tierarztl Prax,1996,24(3):316-8.
[78]张仁舟.犬细小病毒分离鉴定及遗传进化分析[D];中国人民解放军军事医学科学院,2010.
[79]SIMPSON A A, CHANDRASEKAR V, HEBERT B, et al. Host range andvariability of calcium binding by surface loops in the capsids of canine and felineparvoviruses [J]. J Mol Biol,2000,300(3):597-610.
[80]PARRISH C R. Host range relationships and the evolution of canine parvovirus[J]. Vet Microbiol,1999,69(1-2):29-40.
[81]SPITZER A L, PARRISH C R, MAXWELL I H. Tropic determinant for canineparvovirus and feline panleukopenia virus functions through the capsid protein VP2[J]. J Gen Virol,1997,78(Pt4)(925-8.
[82]WEICHERT W S, PARKER J S, WAHID A T, et al. Assaying for structuralvariation in the parvovirus capsid and its role in infection [J]. Virology,1998,250(1):106-17.
[83]CURETON D K, HARBISON C E, COCUCCI E, et al. Limited transferrinreceptor clustering allows rapid diffusion of canine parvovirus into clathrin endocyticstructures [J]. J Virol,2012.
[84]PARKER J S, MURPHY W J, WANG D, et al. Canine and feline parvovirusescan use human or feline transferrin receptors to bind, enter, and infect cells [J]. J Virol,2001,75(8):3896-902.
[2]周云朵.犬细小病毒的分离鉴定与生物学特性研究[D];华中农业大学,2011.
[3] CHINCHKAR S R, MOHANA SUBRAMANIAN B, HANUMANTHA RAO N,et al. Analysis of VP2gene sequences of canine parvovirus isolates in India [J]. ArchVirol,2006,151(9):1881-7.
[4] DOKI M, FUJITA K, MIURA R, et al. Sequence analysis of VP2gene of canineparvovirus isolated from domestic dogs in Japan in1999and2000[J]. CompImmunol Microbiol Infect Dis,2006,29(4):199-206.
[5] MCKENNA R, OLSON N H, CHIPMAN P R, et al. Three-dimensional structureof Aleutian mink disease parvovirus: implications for disease pathogenicity [J]. JVirol,1999,73(8):6882-91.
[6] FILIPOV C, DECARO N, DESARIO C, et al. Canine parvovirus epidemiologyin Bulgaria [J]. J Vet Diagn Invest,2011,23(1):152-4.
[7] ZHAO J, YAN X, WU W.[Origin and evolution of canine parvovirus--a review][J]. Wei Sheng Wu Xue Bao,2011,51(7):869-75.
[8] MOCHIZUKI M, OHSHIMA T, UNE Y, et al. Recombination between vaccineand field strains of canine parvovirus is revealed by isolation of virus in canine andfeline cell cultures [J]. J Vet Med Sci,2008,70(12):1305-14.
[9] LARSON L J, SCHULTZ R D. Do two current canine parvovirus type2and2bvaccines provide protection against the new type2c variant?[J]. Vet Ther,2008,9(2):94-101.
[10]JEOUNG S Y, AHN S J, KIM D.Genetic analysis of VP2gene of canineparvovirus isolates in Korea [J]. J Vet Med Sci,2008,70(7):719-22.
[11] DECARO N,DESARIO C,ADDIE D D, et al. The study molecular epidemiologyof canine parvovirus, Europe [J]. Emerg Infect Dis,2007,13(8):1222-4.
[12]DECARO N, DESARIO C, PARISI A,et al. Genetic analysis of canine parvovirustype2c [J]. Virology,2009,385(1):5-10.
[13]KANG B K, SONG D S, LEE C S, et al. Prevalence and genetic characterizationof canine parvoviruses in Korea [J]. Virus Genes,2008,36(1):127-33.
[14]DECARO N, DESARIO C, ELIA G, et al. Evidence for immunisation failure invaccinated adult dogs infected with canine parvovirus type2c [J]. New Microbiol,2008,31(1):125-30.
[15]CAVALLI A, MARTELLA V, DESARIO C, et al. Evaluation of the antigenicrelationships among canine parvovirus type2variants [J]. Clin Vaccine Immunol,2008,15(3):534-9.
[16]JOAO VIEIRA M, SILVA E, OLIVEIRA J, et al. Canine parvovirus2c infectionin central Portugal [J]. J Vet Diagn Invest,2008,20(4):488-91.
[17]SIEDEK E M, SCHMIDT H, STURE G H, et al. Vaccination with canineparvovirus type2(CPV-2) protects against challenge with virulent CPV-2b andCPV-2c [J]. Berl Munch Tierarztl Wochenschr,2011,124(1-2):58-64.
[18]TOUIHRI L, BOUZID I, DAOUD R, et al. Molecular characterization of canineparvovirus-2variants circulating in Tunisia [J]. Virus Genes,2009,38(2):249-58.
[19]HARBISON C E, LYI S M, WEICHERT W S, et al. Early steps in cell infectionby parvoviruses: host-specific differences in cell receptor binding but similarendosomal trafficking [J]. J Virol,2009,83(20):10504-14.
[20]OHSHIMA T, MOCHIZUKI M. Evidence for recombination between felinepanleukopenia virus and canine parvovirus type2[J]. J Vet Med Sci,2009,71(4):403-8.
[21]VIHINEN-RANTA M, KALELA A, MAKINEN P, et al. Intracellular route ofcanine parvovirus entry [J]. J Virol,1998,72(1):802-6.
[22]PARKER J S, PARRISH C R. Canine parvovirus host range is determined by thespecific conformation of an additional region of the capsid [J]. J Virol,1997,71(12):9214-22.
[23]HOELZER K, SHACKELTON L A, PARRISH C R,et al. Phylogenetic analysisreveals the emergence, evolution and dispersal of carnivore parvoviruses [J]. J GenVirol,2008,89(Pt9):2280-9.
[24]CALDERON M G,ROMANUTTI C, A D A,et al. Evolution of canine parvovirusin Argentina between years2003and2010: CPV2c has become the predominantvariant affecting the domestic dog population [J]. Virus Res,2011,157(1):106-10.
[25]贺英,曹旺斌,史秋梅等.基于转铁蛋白受体对犬细小病毒原代细胞组织嗜性的研究,中国黑龙江齐齐哈尔, F,2010[C].
[26]PARKER J S, PARRISH C R. Cellular uptake and infection by canine parvovirusinvolves rapid dynamin-regulated clathrin-mediated endocytosis, followed by slowerintracellular trafficking [J]. J Virol,2000,74(4):1919-30.
[27]耿志贤,时彦胜,郑亚平等.犬细小病毒病的流行现状调查[J].中国畜牧兽医,2009,11):135-7.
[28]BATTILANI M, BALBONI A, USTULIN M, et al. Genetic complexity andmultiple infections with more Parvovirus species in naturally infected cats [J]. Vet Res,2011,42(1):43.
[29]VIHINEN-RANTA M, YUAN W, PARRISH C R. Cytoplasmic trafficking of thecanine parvovirus capsid and its role in infection and nuclear transport [J]. J Virol,2000,74(10):4853-9.
[30]NTAFIS V, XYLOURI E, KALLI I, et al. Characterization of Canine parvovirus2variants circulating in Greece [J]. J Vet Diagn Invest,2010,22(5):737-40.
[31]MEERS J, KYAW-TANNER M, BENSINK Z, et al. Genetic analysis of canineparvovirus from dogs in Australia [J]. Aust Vet J,2007,85(10):392-6.
[32]WU Z, SUN H, ZHANG X.[Immunoadjuvant effect of the extracellular domainof canine CTLA-4on the VP2protein of canine parvovirus][J]. Wei Sheng Wu XueBao,2008,48(3):369-74.
[33]HAFENSTEIN S, BOWMAN V D, SUN T, et al. Structural comparison ofdifferent antibodies interacting with parvovirus capsids [J]. J Virol,2009,83(11):5556-66.
[34]CASTRO T X, COSTA E M, LEITE J P, et al. Monitoring of canine parvovirus(CPV) strains detected in vaccinated puppies in Brazil [J]. Res Vet Sci,2011,90(2):336-40.
[35]TAGUCHI M, NAMIKAWA K, MARUO T, et al. Antibody titers for canineparvovirus type-2, canine distemper virus, and canine adenovirus type-1in adulthousehold dogs [J]. Can Vet J,2011,52(9):983-6.
[36]贺英,张曼夫,曹旺斌.犬细小病毒宿主范围和入侵途径研究进展[J].畜牧与兽医,2009,01):101-4.
[37]SPIBEY N, GREENWOOD N M, SUTTON D, et al. Canine parvovirus type2vaccine protects against virulent challenge with type2c virus [J]. Vet Microbiol,2008,128(1-2):48-55.
[38]MARULAPPA S Y, KAPIL S. Simple tests for rapid detection of canineparvovirus antigen and canine parvovirus-specific antibodies [J]. Clin VaccineImmunol,2009,16(1):127-31.
[39]ELIA G, CAVALLI A, DESARIO C, et al. Detection of infectious canineparvovirus type2by mRNA real-time RT-PCR [J]. J Virol Methods,2007,146(1-2):202-8.
[40]宋永奇,李玉燕,吕艳丽等.3种犬细小病毒感染诊断方法比较[J].畜牧与兽医,2008,06):31-5.
[41]郭伟.犬细小病毒分离鉴定及环介导等温扩增快速检测方法的建立[D];内蒙古农业大学,2010.
[42]GODSALL S A, CLEGG S R, STAVISKY J H, et al. Epidemiology of canineparvovirus and coronavirus in dogs presented with severe diarrhoea to PDSA PetAidhospitals [J]. Vet Rec,2010,167(6):196-201.
[43]CHO H S, KANG J I, PARK N Y. Detection of canine parvovirus in fecalsamples using loop-mediated isothermal amplification [J]. J Vet Diagn Invest,2006,18(1):81-4.
[44]DECARO N, MARTELLA V, DESARIO C, et al. First detection of canineparvovirus type2c in pups with haemorrhagic enteritis in Spain [J]. J Vet Med BInfect Dis Vet Public Health,2006,53(10):468-72.
[45]杨颗粒,张丰范,孙玉彬.犬细小病毒病的诊断和治疗[J].农村科技,1995,07):21.
[46]CLEGG S R, COYNE K P, PARKER J, et al. Molecular epidemiology andphylogeny reveal complex spatial dynamics in areas where canine parvovirus isendemic [J]. J Virol,2011,85(15):7892-9.
[47]BRAGG R F, DUFFY A L, DECECCO F A, et al. Clinical evaluation of a singledose of immune plasma for treatment of canine parvovirus infection [J]. J Am VetMed Assoc,2012,240(6):700-4.
[48]孙艳芳,贾宁,扎西英派等.犬细小病毒病的病理学观察; proceedings of the中国畜牧兽医学会兽医病理学分会第十五次、中国病理生理学会动物病生专业委员会第十四次学术讨论会,中国黑龙江哈尔滨, F,2007[C].
[49]ZHANG R, YANG S, ZHANG W, et al. Phylogenetic analysis of the VP2gene ofcanine parvoviruses circulating in China [J]. Virus Genes,2010,40(3):397-402.
[50]MAJER-DZIEDZIC B, JAKUBCZAK A, ZIETEK J. Phylogenetic analysis ofcanine parvovirus CPV-2strains and its variants isolated in Poland [J]. Pol J Vet Sci,2011,14(3):379-84.
[51]任宝波,孙冰.犬细小病毒感染防治的研究进展[J].养殖技术顾问,2007,04):60-1.
[52]YOON S H, JEONG W, KIM H J, et al. Molecular insights into the phylogeny ofcanine parvovirus2(CPV-2) with emphasis on Korean isolates: a Bayesian approach[J]. Arch Virol,2009,154(8):1353-60.
[53]陈琛,邬静,张海亮等.犬细小病毒病诊断与防治情况调查[J].湖北农业科学,2010,08):1926-8.
[54]PRATELLI A, CAVALLI A, NORMANNO G, et al. Immunization of pups withmaternally derived antibodies to canine parvovirus (CPV) using a modified-livevariant (CPV-2b)[J]. J Vet Med B Infect Dis Vet Public Health,2000,47(4):273-6.
[55]王洪林.犬细小病毒北京株分离和动物发病模型建立[D];中国农业科学院,2009.
[56]JIANG W, BAKER H J, SWANGO L J, et al. Nucleic acid immunization protectsdogs against challenge with virulent canine parvovirus [J]. Vaccine,1998,16(6):601-7.
[57]MOHAN RAJ J, MUKHOPADHYAY H K, THANISLASS J, et al. Isolation,molecular characterization and phylogenetic analysis of canine parvovirus [J]. InfectGenet Evol,2010,10(8):1237-41.
[58]DECARO N, BUONAVOGLIA C. Canine parvovirus--a review ofepidemiological and diagnostic aspects, with emphasis on type2c [J]. Vet Microbiol,2012,155(1):1-12.
[59]李晋涛,魏静,牟芝蓉等.人源轮状病毒的小型猪腹泻模型建立[J].中国比较医学杂志,2006,09):577.
[60]廖列如,张林,孔庆波等.犬冠状病毒的分离及鉴定[J].安徽农业科学,2007,30):9549-51+98.
[61]LITSTER A, NICHOLS J, VOLPE A. Prevalence of positive antibody test resultsfor canine parvovirus (CPV) and canine distemper virus (CDV) and response tomodified live vaccination against CPV and CDV in dogs entering animal shelters [J].Vet Microbiol,2011,
[62]邓朝晖,许春娣,陈舜年.新生小鼠轮状病毒腹泻模型建立及病理学致病机制研究[J].临床儿科杂志,2008,10):862-5.
[63]LARSON L J, SCHULTZ R D. Three-year serologic immunity against canineparvovirus type2and canine adenovirus type2in dogs vaccinated with a caninecombination vaccine [J]. Vet Ther,2007,8(4):305-10.
[64]MOON H S, LEE S A, LEE S G, et al. Comparison of the pathogenicity in threedifferent Korean canine parvovirus2(CPV-2) isolates [J]. Vet Microbiol,2008,131(1-2):47-56.
[65]阿拉坦布日古德,刘晓松,张喜彦等.犬细小病毒实验感染研究[J].内蒙古兽医,1995,02):1-4.
[66]IRIS K, LEONTIDES L S, MYLONAKIS M E, et al. Factors affecting theoccurrence, duration of hospitalization and final outcome in canine parvovirusinfection [J]. Res Vet Sci,2010,89(2):174-8.
[67]GOODMAN L B, LYI S M, JOHNSON N C, et al. Binding site on the transferrinreceptor for the parvovirus capsid and effects of altered affinity on cell uptake andinfection [J]. J Virol,2010,84(10):4969-78.
[68]NYKKY J, TUUSA J E, KIRJAVAINEN S, et al. Mechanisms of cell death incanine parvovirus-infected cells provide intuitive insights to developing nanotools formedicine [J]. Int J Nanomedicine,2010,5(417-28.
[69]MUZ D, OGUZOGLU T C, TIMURKAN M O, et al. Characterization of thepartial VP2gene region of canine parvoviruses in domestic cats from Turkey [J].Virus Genes,2012,44(2):301-8.
[70]GALLO CALDERON M, WILDA M, BOADO L, et al. Study of canineparvovirus evolution: comparative analysis of full-length VP2gene sequences fromArgentina and international field strains [J]. Virus Genes,2012,44(1):32-9.
[71]LLAMAS-SAIZ A L, AGBANDJE-MCKENNA M, PARKER J S, et al.Structural analysis of a mutation in canine parvovirus which controls antigenicity andhost range [J]. Virology,1996,225(1):65-71.
[72]CHANG S F, SGRO J Y, PARRISH C R. Multiple amino acids in the capsidstructure of canine parvovirus coordinately determine the canine host range andspecific antigenic and hemagglutination properties [J]. J Virol,1992,66(12):6858-67.
[73]GEMMA T, KAI C, MIKAMI T. Suppression of canine parvovirus growth inCRFK cells by canine distemper virus [J]. J Vet Med Sci,1995,57(3):535-7.
[74]CHAPMAN M S, ROSSMANN M G. Structure, sequence, and functioncorrelations among parvoviruses [J]. Virology,1993,194(2):491-508.
[75]PARRISH C R. Pathogenesis of feline panleukopenia virus and canine parvovirus[J]. Baillieres Clin Haematol,1995,8(1):57-71.
[76]TRUYEN U.[Canine parvovirus: recent knowledge of the origin anddevelopment of a viral pathogen][J]. Tierarztl Prax,1994,22(6):579-84.
[77]TRUYEN U.[Evolution of canine parvovirus: loss and gain of the feline host][J].Tierarztl Prax,1996,24(3):316-8.
[78]张仁舟.犬细小病毒分离鉴定及遗传进化分析[D];中国人民解放军军事医学科学院,2010.
[79]SIMPSON A A, CHANDRASEKAR V, HEBERT B, et al. Host range andvariability of calcium binding by surface loops in the capsids of canine and felineparvoviruses [J]. J Mol Biol,2000,300(3):597-610.
[80]PARRISH C R. Host range relationships and the evolution of canine parvovirus[J]. Vet Microbiol,1999,69(1-2):29-40.
[81]SPITZER A L, PARRISH C R, MAXWELL I H. Tropic determinant for canineparvovirus and feline panleukopenia virus functions through the capsid protein VP2[J]. J Gen Virol,1997,78(Pt4)(925-8.
[82]WEICHERT W S, PARKER J S, WAHID A T, et al. Assaying for structuralvariation in the parvovirus capsid and its role in infection [J]. Virology,1998,250(1):106-17.
[83]CURETON D K, HARBISON C E, COCUCCI E, et al. Limited transferrinreceptor clustering allows rapid diffusion of canine parvovirus into clathrin endocyticstructures [J]. J Virol,2012.
[84]PARKER J S, MURPHY W J, WANG D, et al. Canine and feline parvovirusescan use human or feline transferrin receptors to bind, enter, and infect cells [J]. J Virol,2001,75(8):3896-902.