新疆蚊传虫媒病毒调查
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摘要
新疆位于中国西北边陲,是中国面积最大的省级行政区。新疆陆地边境线长,周边与俄罗斯、印度、蒙古等八个国家接壤。自北向南,阿勒泰山脉、天山山脉和昆仑山脉将新疆分隔成北疆和南疆,使新疆在气候、地理、生境和物种等方面存在多样性并与内地差异明显。新疆蚊虫媒介种类较多,与人畜关系密切。近年来在新疆进行的虫媒病毒调查中从蚊和蜱中分离到辛德毕斯、东方马脑炎、西方马脑炎等多种虫媒病毒。然而,这些病毒多分离自气候温润、蚊虫密度较大的北疆,南疆地区虫媒病毒调查情况少有报道。然而,南疆地区病毒性脑炎流行病学监测结果显示,在蚊虫活动高峰的夏秋季节,当地存在病因不明的发热和病毒性脑炎的流行。然而,我国其他地区流行的在夏季引起病毒性脑炎的乙脑病毒在新疆尚未发现。这些信息提示当地流行的虫媒病毒及相关疾病可能与内地存在较大差异。因此,对新疆地区进行系统的虫媒病毒病原学的调查及相关病毒的人群感染的血清流行病学研究对于预防和控制当地虫媒病毒病均具有重要意义。
本课题分别对我国新疆南,北疆地区连续3年开展虫媒病毒调查,在采集的蚊虫标本中分离到大量虫媒病毒,其中首次在我国分离到布尼亚病毒科Tahyna病毒,并证明当地不明原因发热患者中存在该病毒IgM抗体阳性,文章发表在Emerg Infect Dis.2009,15(2):306-309。
1.新疆蚊传虫媒病毒调查
2006年、2007年和2008年7～8月在新疆地区喀什地区的伽师县、和田地区的民丰县、巴音郭楞蒙古自治州的尉犁县和博湖县、伊犁哈萨克自治州的伊宁县、阿勒泰地区的阿勒泰市和布尔津县、塔城地区的沙湾县以及哈密地区哈密市采集蚊虫标本21405只,绝大多数为背点伊蚊标本,分222批进行研磨和捕痉掷搿＠米橹嘌赴ǚ掷氩《?结果分离到50株阳性分离物,均来自背点伊蚊标本。经过系统鉴定。共得到3株Tahyna病毒(Tahyna virus,TAHV)、41株辽宁病毒(Liaoning virus,LNV)和6株待鉴定阳性分离物。
2.新疆新分离病毒的分子生物学特征研究
本次研究对分离自喀什地区伽师县,巴音郭楞蒙古自治州的尉犁县的3株TAHV进行序列测定和分子特征分析,测定了这3株(XJ30625、XJ0708和XJ0710)的全基因组序列(L、M和S节段编码区序列)。对TAHV病毒系统进化分析和核苷酸与氨基酸差异分析结果显示,TAHV各节段中S节段编码区同源性最商,中国株之间核苷酸同源性99.2-99.6%,氨基酸同源性为100%,与国外毒株相比,核苷酸同源性为92.2-92.9%,氨基酸同源性均为98.3%;L节段序列分析显示,中国株之间核苷酸同源性为96.3-97.4%,氨基酸同源性均为100%,与国外毒株相比核苷酸同源性81-82.5%,氨基酸同源性97.8%,提示多为同义突变:对于病毒M节段,我国新疆分离株之间核苷酸同源性80.7-96.1%,氨基酸同源性92.6-99%:与国外分离株核苷酸同源性80.1-82.1%,氨基酸同源性92.6-94.2%。S和L节段中国株之间各节段同源性较高,而与欧洲流行株Bardos 92同源性较低,体现了病毒的地域分布特征。M节段中国株之间存在较大变异,提示中国分离株可能存在多个亚型。结合噬斑滴定试验结果,推测这一节段的变异可能会影响TAHV噬斑形态及病毒生物学行为。
本次试验对41株分离自喀什地区、巴音郭楞蒙古自治州和阿勒泰地区的LNV的第10和第12节段的进行序列测定、系统进化分析和核苷酸、氨基酸差异分析。结果显示,新疆地区分离的LNV毒株同源性较高,第十节段核苷酸同源性均大于94%,而与该病毒东北原型株核苷酸同源性仅75%左右。依据第10节段核苷酸序列信息可以将LNV分成四个亚型。南疆地区LNV属于基因Ⅰ型,而北疆阿勒泰地区分离到Ⅰ型和Ⅱ型,其中Ⅱ型LNV与东北地区原型株Ⅲ和Ⅳ型存在着许多特征性核苷酸改变,说明Ⅱ型和Ⅲ、Ⅳ型病毒之间可能存在传播关系。并且,推测在这两个地区之间的省区同样存在LNV流行的可能。而且,流行的病毒型别应该介于Ⅱ型和Ⅲ型之间。
本研究对蚊虫标本中分离的6株引起C6/36细胞病变的病毒分离物进行了鉴定,结果显示,分离物对多种虫媒病毒组特异性免疫血清不产生反应,应用甲病毒、黄病毒、布尼亚病毒以及呼肠孤(环状病毒、Seadonavirus等)病毒属特异性引物的基因扩增反应也呈阴性,提示新分离病毒可能不属于以上病毒属,进一步的鉴定工作正在进行中。
3.新疆夏季不明原因发热及脑炎患者虫媒病毒感染调查
为了明确TAHV与当地人群疾病的关系,本研究连续2年在新疆采集不明原因发热病人急性期血清标本964例,应用IFA方法进行血清TAHV感染调查。其中,喀什地区采集不明原因发热患者742例,检测IgM抗体阳性率5.26%(39/742),IgG抗体阳性率18.27%(59/323);在北疆地区的伊犁哈萨克自治州察布查尔锡伯自治县采集不明原因发热患者标本222例,并未发现TAHV IgM抗体阳性者。这些结果提示,新疆南疆喀什地区不明原因发热人群中存在该病毒的感染。结合TAHV病毒分离结果,认为南疆多个地区存在TAHV循环及人群感染。
对2007年伽师县采集的10例TAHV IgM抗体阳性血清及恢复期血清进行中和试验。结果发现,所有标本中和抗体指数介于1:10-1:80之间,提示当地人群中存在该病毒的感染,但是未能发现恢复期较急性期血清抗体呈现4倍增长,因此尚不能证实TAHV是这些IgM抗体阳性病例发热疾病的病因。本研究还发现,应用IFA检测血清TAHV抗体时所得到的抗体滴度与中和试验检测时得到的抗体滴度基本相符,提示IFA方法可以用来进行TAHV病毒抗体的初筛。
本研究还对新疆南疆伽师县2004年夏季发生的一起乙型脑炎流行的病因开展了进一步研究。结果显示疫情期间采集的6对急性期和恢复期血清标本的乙型脑炎病毒(JEV)IgM抗体均呈阳性,但是只有两对标本存在低滴度JEV中和抗体4倍增高(5:20和5:40),其余4对标本JEV中和抗体呈阴性反应,根据此结果认为该起疫情为乙脑流行尚显证据不足。对以上标本开展了多种虫媒病毒抗体筛查,结果发现这些标本JEV、登革病毒(DENV)、西尼罗病毒(WNV)、圣路易斯脑炎病毒(SLEV)等多种黄病毒IgM和IgG抗体均呈现阳性(TAHV及其它甲病毒属虫媒病毒抗体阴性),考虑到黄病毒之间存在血清学交叉反应,这些结果仅提示当地可能存在某种黄病毒的感染,但对其真正病因尚无法判断。建议今后有条件时开展多种黄病毒中和试验以进一步确定病因。
此外,本研究对2008年新疆南北疆地区采集的641例不明原因发热患者血清进行了WNV、DENV和JEV虫媒病毒IgM抗体的筛查。结果发现11例患者血清WNV抗体阳性,19例患者血清DENV抗体阳性和4例患者血清JEV抗体阳性。这些结果提示,新疆地区存在多种蚊传黄病毒属虫媒病毒,并有可能引起夏季人群不明原因发热等疾病。
综上所述,本研究利用3年时间在我国新疆地区开展了虫媒病毒系统调查,分离到多种虫媒病毒,首次证明新疆南疆地区存在Tahyna病毒感染,这些数据不但丰富了我国和新疆维吾尔自治区虫媒病毒的种类,而且为了解当地传染病病因以及积极预防和控制相关虫媒病毒疾病提供了重要的信息。进一步工作应该侧重于新疆当地及全国范围内Tahyna病毒感染的病例发现,临床表现,疾病负担以及该病在新疆以及我国其他地区的流行范围等。
Xinjaing is the largest provincial-level administrative region in China.It is adjacent to five provinces in China and borders on eight countries.Xinjiang is divided into North and South Regions by Tianshan Mountain which lies across the middle of Xinjiang.There are remarkable differences on aspects of climate,geography, environment and species distribution between North and South Regions of Xinjiang. Many species of arthropods are distributed in Xinjiang.Mosquitoe is major vector, which might transmit arboviruses and cause zoonotic diseases.Recent years several arboviruses such as Sindbis,Eastern equine encephalitis,Western equine encephalitis have been isolated from Xinjiang.But until now there were still no reports of population-related diseases.However,the viral encephalitis epidemiological surveillance in Xinjiang showed that there were unknown fever and viral encephalitis cases during summer.In addition,most provinces report Japanese encephalitis cases during summer except in Xinjiang.The information showed that there is significant difference between Xinjiang and other parts of China.So we carried out a systematic investigation of arbovirus etiology and correlated human infection with seroepidemiological studies.
In this stydy,system investigation of arbovirus was carried out to clearly describe the existence of the species of arbovirus and distribution.Further study also identify the taxonomy and characteristics of molecular biology of the newly isolates. Finally,correlation between the arbovirus and human illness was stydied.
1.Arbovirus investigation in Xinjiang
Mosquito samples were collected in the villages of 11 counties belonged to Kashi, Hetian,Bayinguoleng,Aletai,Yili,Tacheng,Hami,from July to Septrmber in 2006, 2007 and 2008.21405 individuals had been collected.The mosquitoes were sorted into 222 pools for virus isolation.Total 50 virus isolates were obtained.47 isolates were observed to cause CPE in C6/36 cells only,while the other 3 isolates could cause CPE in both C6/36 cells and BHK-21 cells.All the isolates were identified by using morphological,serological and molecular biological methods.They are 3 Tahyna viruses(TAHV),41 Liaoning viruses,and 6 strains of unidentified virus.
2.Molecular analysis of the new isolates
In this study 3 strains of TAHV isolated from Kashi and Bayinguoleng area were sequenced.Phylogenetic analysis and differences of nucleotides and amino acids were carried out.The results showed that S segment had the highest homology among all the 3 segments,which were 98.3%-100%for nucleotide and 98.3%-100%for amino acids respectively,all the Chinese strains are between 99.2-99.6%for nucleotide and100%for amino acid.There are 81-97.4%and 97.8-100%similarity for L segment. M segment is the lowest similiarity one among all 3 segments.All these 3 Chinese strains showed higher identity than Europe strains.So it was concluded that there are correlation between the geographical distribution and the genomic character.
41 strains of LNV were isolated from Kashi,Bayinguoleng and Aletai region. Segment 10 and segment 12 of LNV were sequenced.The phylogenetic analysis showed that LNV isolates from Xinjiang showed high similiarity than Jilin isolates. The identities of the 10~(th) segment between all Xinjiang strains were more than 94%in nucleotide and only 75%with Jilin strains.4 subtypes of LNV were classified based on nucleotide sequence of segment 10.All strains from southern region of Xinjiang belong to genotypeⅠ.GenotypeⅠandⅡhave been isolated from Aletai.Strains from Jilin were classified as genotypeⅢandⅣ.GenotypeⅡ,ⅢandⅣhave some consistency mutation.So correlation may exist between all these subtypes.
3.Arbovirus and correlation with illness among the local population in Xinjiang.
Among them,742 cases were collected from Kashi,IgM positive rate was 5.26% (39/742),and IgG positive rate was 18.27%(59/323),no IgM positive were found in northern part of Xinjiang among 222 cases in Yili.The results indicated that in southern part of Xinjiang there were TAHV cycle and local people may be infected during Sumer and for northern part of Xinjiang,due to the Tianshan Mountain,TAHV may be not so active and few people were infected with it.
To determine whether the initial illness of the patients was correlated with TAHV infection,in December 2007 second serum samples from 10 of the IgM positive patients.These paired serum specimens were tested by serum dilution neutralization test with XJ0625 virus.Most of the samples contained neutralizing antibody with a titer of 40 or 80.The results indicated that human infection with TAHV is common in the area.Furthermore,the result of IFA test coordinate with the NT test,so it may be used as a preliminary screening test.
Sera samples of acute and convalescent phase of unknown encephalitis patients were screened by IFA test for several kinds of arbovirus.The results showed all the sera samples have IgG and IgM against flavivirus with a high titer and no positive results for non-flavirus.Combination with the features of clinical manifestations,prognostic and immunological responsds,it was concluded that pathogen of the epidemic may be a kind of unknown flavivirus.
In the study,an pathogen investigation on an encephalitis outbreak in south Xinjiang in the summer of 2004 was conducted.The results showed only 2cases had 4 fold rises among all six pairs of acute and convalescent serum specimens collected during the disease.And the neutralizing antibody titer against JEV were very low.The remaining four pairs of specimens of JEV neutralizing antibodies were negative, according to the results,it was not sufficient to take JEV as the pathogen for the endemic.So,a wide range of arbovirus antibody screening was carried out.The results showed that IgM and IgG antibody against JEV,DENV,WNV,SLEV and other flavirus were positive and negative for TAHV and other alphavirus.Taking into account serological cross-reaction between flavivirus,these results prompted only there may be some flavivirus infection.But the real pathogen is still not certain.It is recommended that NT test against a variety of flavivirus should be carried out.
In addition,641 cases of patients with unknown fever in northern and southern Xinjiang were collected in 2008 to carried out the IgM antibody screening against WNV,DENV and JEV.The results showed that 11 cases has IgM antibody against WNV,19 cases against DENV and 4 cases against JEV.These results indicated the existence of a wide range of the flavivirus in Xinjiang and may cause human diseases in summer.
The importance of the study
In summary,arbovirus investigation in Xinjang was carried out in this study during 2006,2007 and 2008.3 strains of TAHV and 41 strains of LNV were isolated from mosquitoe pools.The isolation of TAHV from southern region of Xinjiang indicated this virus prevelant in local area and no isolation from northern region of Xinjiang indicated that this virus may not active there.Molecular character of all the viruses indicated that there may be several subtypes existed in Xinjiang. Seroepidemiological results showed TAHV may be prevelant in local area and unknown fever patients may be infected by such virus.
LNV existed in most region of Xinjiang,the sequence of all the strains were relatively conserved.Phylogenetic results showed four subtypes can be classified based on segment 10.Isolates from the Xinjiang region,and Jilin exist significant regional comparison.Isolates from Aletai showed relatively high homology and the characteristic consensus sequences with Jilin isolates.It suggested that there may be similar virus between Jilin and Xinjiang.
These findings give us a clue to strengthen arbovirus surveillance in order to prevent arbovirus diseases.Identification and molecular analysis of TAHV and LNV isolates isolated from Xinjiang can help to know more about the virus characterization, especially the molecular difference from foreign isolates,and can provide basic information for further study of the arboviruses distributed in Chinese territory.
本课题分别对我国新疆南,北疆地区连续3年开展虫媒病毒调查,在采集的蚊虫标本中分离到大量虫媒病毒,其中首次在我国分离到布尼亚病毒科Tahyna病毒,并证明当地不明原因发热患者中存在该病毒IgM抗体阳性,文章发表在Emerg Infect Dis.2009,15(2):306-309。
1.新疆蚊传虫媒病毒调查
2006年、2007年和2008年7～8月在新疆地区喀什地区的伽师县、和田地区的民丰县、巴音郭楞蒙古自治州的尉犁县和博湖县、伊犁哈萨克自治州的伊宁县、阿勒泰地区的阿勒泰市和布尔津县、塔城地区的沙湾县以及哈密地区哈密市采集蚊虫标本21405只,绝大多数为背点伊蚊标本,分222批进行研磨和捕痉掷搿＠米橹嘌赴ǚ掷氩《?结果分离到50株阳性分离物,均来自背点伊蚊标本。经过系统鉴定。共得到3株Tahyna病毒(Tahyna virus,TAHV)、41株辽宁病毒(Liaoning virus,LNV)和6株待鉴定阳性分离物。
2.新疆新分离病毒的分子生物学特征研究
本次研究对分离自喀什地区伽师县,巴音郭楞蒙古自治州的尉犁县的3株TAHV进行序列测定和分子特征分析,测定了这3株(XJ30625、XJ0708和XJ0710)的全基因组序列(L、M和S节段编码区序列)。对TAHV病毒系统进化分析和核苷酸与氨基酸差异分析结果显示,TAHV各节段中S节段编码区同源性最商,中国株之间核苷酸同源性99.2-99.6%,氨基酸同源性为100%,与国外毒株相比,核苷酸同源性为92.2-92.9%,氨基酸同源性均为98.3%;L节段序列分析显示,中国株之间核苷酸同源性为96.3-97.4%,氨基酸同源性均为100%,与国外毒株相比核苷酸同源性81-82.5%,氨基酸同源性97.8%,提示多为同义突变:对于病毒M节段,我国新疆分离株之间核苷酸同源性80.7-96.1%,氨基酸同源性92.6-99%:与国外分离株核苷酸同源性80.1-82.1%,氨基酸同源性92.6-94.2%。S和L节段中国株之间各节段同源性较高,而与欧洲流行株Bardos 92同源性较低,体现了病毒的地域分布特征。M节段中国株之间存在较大变异,提示中国分离株可能存在多个亚型。结合噬斑滴定试验结果,推测这一节段的变异可能会影响TAHV噬斑形态及病毒生物学行为。
本次试验对41株分离自喀什地区、巴音郭楞蒙古自治州和阿勒泰地区的LNV的第10和第12节段的进行序列测定、系统进化分析和核苷酸、氨基酸差异分析。结果显示,新疆地区分离的LNV毒株同源性较高,第十节段核苷酸同源性均大于94%,而与该病毒东北原型株核苷酸同源性仅75%左右。依据第10节段核苷酸序列信息可以将LNV分成四个亚型。南疆地区LNV属于基因Ⅰ型,而北疆阿勒泰地区分离到Ⅰ型和Ⅱ型,其中Ⅱ型LNV与东北地区原型株Ⅲ和Ⅳ型存在着许多特征性核苷酸改变,说明Ⅱ型和Ⅲ、Ⅳ型病毒之间可能存在传播关系。并且,推测在这两个地区之间的省区同样存在LNV流行的可能。而且,流行的病毒型别应该介于Ⅱ型和Ⅲ型之间。
本研究对蚊虫标本中分离的6株引起C6/36细胞病变的病毒分离物进行了鉴定,结果显示,分离物对多种虫媒病毒组特异性免疫血清不产生反应,应用甲病毒、黄病毒、布尼亚病毒以及呼肠孤(环状病毒、Seadonavirus等)病毒属特异性引物的基因扩增反应也呈阴性,提示新分离病毒可能不属于以上病毒属,进一步的鉴定工作正在进行中。
3.新疆夏季不明原因发热及脑炎患者虫媒病毒感染调查
为了明确TAHV与当地人群疾病的关系,本研究连续2年在新疆采集不明原因发热病人急性期血清标本964例,应用IFA方法进行血清TAHV感染调查。其中,喀什地区采集不明原因发热患者742例,检测IgM抗体阳性率5.26%(39/742),IgG抗体阳性率18.27%(59/323);在北疆地区的伊犁哈萨克自治州察布查尔锡伯自治县采集不明原因发热患者标本222例,并未发现TAHV IgM抗体阳性者。这些结果提示,新疆南疆喀什地区不明原因发热人群中存在该病毒的感染。结合TAHV病毒分离结果,认为南疆多个地区存在TAHV循环及人群感染。
对2007年伽师县采集的10例TAHV IgM抗体阳性血清及恢复期血清进行中和试验。结果发现,所有标本中和抗体指数介于1:10-1:80之间,提示当地人群中存在该病毒的感染,但是未能发现恢复期较急性期血清抗体呈现4倍增长,因此尚不能证实TAHV是这些IgM抗体阳性病例发热疾病的病因。本研究还发现,应用IFA检测血清TAHV抗体时所得到的抗体滴度与中和试验检测时得到的抗体滴度基本相符,提示IFA方法可以用来进行TAHV病毒抗体的初筛。
本研究还对新疆南疆伽师县2004年夏季发生的一起乙型脑炎流行的病因开展了进一步研究。结果显示疫情期间采集的6对急性期和恢复期血清标本的乙型脑炎病毒(JEV)IgM抗体均呈阳性,但是只有两对标本存在低滴度JEV中和抗体4倍增高(5:20和5:40),其余4对标本JEV中和抗体呈阴性反应,根据此结果认为该起疫情为乙脑流行尚显证据不足。对以上标本开展了多种虫媒病毒抗体筛查,结果发现这些标本JEV、登革病毒(DENV)、西尼罗病毒(WNV)、圣路易斯脑炎病毒(SLEV)等多种黄病毒IgM和IgG抗体均呈现阳性(TAHV及其它甲病毒属虫媒病毒抗体阴性),考虑到黄病毒之间存在血清学交叉反应,这些结果仅提示当地可能存在某种黄病毒的感染,但对其真正病因尚无法判断。建议今后有条件时开展多种黄病毒中和试验以进一步确定病因。
此外,本研究对2008年新疆南北疆地区采集的641例不明原因发热患者血清进行了WNV、DENV和JEV虫媒病毒IgM抗体的筛查。结果发现11例患者血清WNV抗体阳性,19例患者血清DENV抗体阳性和4例患者血清JEV抗体阳性。这些结果提示,新疆地区存在多种蚊传黄病毒属虫媒病毒,并有可能引起夏季人群不明原因发热等疾病。
综上所述,本研究利用3年时间在我国新疆地区开展了虫媒病毒系统调查,分离到多种虫媒病毒,首次证明新疆南疆地区存在Tahyna病毒感染,这些数据不但丰富了我国和新疆维吾尔自治区虫媒病毒的种类,而且为了解当地传染病病因以及积极预防和控制相关虫媒病毒疾病提供了重要的信息。进一步工作应该侧重于新疆当地及全国范围内Tahyna病毒感染的病例发现,临床表现,疾病负担以及该病在新疆以及我国其他地区的流行范围等。
Xinjaing is the largest provincial-level administrative region in China.It is adjacent to five provinces in China and borders on eight countries.Xinjiang is divided into North and South Regions by Tianshan Mountain which lies across the middle of Xinjiang.There are remarkable differences on aspects of climate,geography, environment and species distribution between North and South Regions of Xinjiang. Many species of arthropods are distributed in Xinjiang.Mosquitoe is major vector, which might transmit arboviruses and cause zoonotic diseases.Recent years several arboviruses such as Sindbis,Eastern equine encephalitis,Western equine encephalitis have been isolated from Xinjiang.But until now there were still no reports of population-related diseases.However,the viral encephalitis epidemiological surveillance in Xinjiang showed that there were unknown fever and viral encephalitis cases during summer.In addition,most provinces report Japanese encephalitis cases during summer except in Xinjiang.The information showed that there is significant difference between Xinjiang and other parts of China.So we carried out a systematic investigation of arbovirus etiology and correlated human infection with seroepidemiological studies.
In this stydy,system investigation of arbovirus was carried out to clearly describe the existence of the species of arbovirus and distribution.Further study also identify the taxonomy and characteristics of molecular biology of the newly isolates. Finally,correlation between the arbovirus and human illness was stydied.
1.Arbovirus investigation in Xinjiang
Mosquito samples were collected in the villages of 11 counties belonged to Kashi, Hetian,Bayinguoleng,Aletai,Yili,Tacheng,Hami,from July to Septrmber in 2006, 2007 and 2008.21405 individuals had been collected.The mosquitoes were sorted into 222 pools for virus isolation.Total 50 virus isolates were obtained.47 isolates were observed to cause CPE in C6/36 cells only,while the other 3 isolates could cause CPE in both C6/36 cells and BHK-21 cells.All the isolates were identified by using morphological,serological and molecular biological methods.They are 3 Tahyna viruses(TAHV),41 Liaoning viruses,and 6 strains of unidentified virus.
2.Molecular analysis of the new isolates
In this study 3 strains of TAHV isolated from Kashi and Bayinguoleng area were sequenced.Phylogenetic analysis and differences of nucleotides and amino acids were carried out.The results showed that S segment had the highest homology among all the 3 segments,which were 98.3%-100%for nucleotide and 98.3%-100%for amino acids respectively,all the Chinese strains are between 99.2-99.6%for nucleotide and100%for amino acid.There are 81-97.4%and 97.8-100%similarity for L segment. M segment is the lowest similiarity one among all 3 segments.All these 3 Chinese strains showed higher identity than Europe strains.So it was concluded that there are correlation between the geographical distribution and the genomic character.
41 strains of LNV were isolated from Kashi,Bayinguoleng and Aletai region. Segment 10 and segment 12 of LNV were sequenced.The phylogenetic analysis showed that LNV isolates from Xinjiang showed high similiarity than Jilin isolates. The identities of the 10~(th) segment between all Xinjiang strains were more than 94%in nucleotide and only 75%with Jilin strains.4 subtypes of LNV were classified based on nucleotide sequence of segment 10.All strains from southern region of Xinjiang belong to genotypeⅠ.GenotypeⅠandⅡhave been isolated from Aletai.Strains from Jilin were classified as genotypeⅢandⅣ.GenotypeⅡ,ⅢandⅣhave some consistency mutation.So correlation may exist between all these subtypes.
3.Arbovirus and correlation with illness among the local population in Xinjiang.
Among them,742 cases were collected from Kashi,IgM positive rate was 5.26% (39/742),and IgG positive rate was 18.27%(59/323),no IgM positive were found in northern part of Xinjiang among 222 cases in Yili.The results indicated that in southern part of Xinjiang there were TAHV cycle and local people may be infected during Sumer and for northern part of Xinjiang,due to the Tianshan Mountain,TAHV may be not so active and few people were infected with it.
To determine whether the initial illness of the patients was correlated with TAHV infection,in December 2007 second serum samples from 10 of the IgM positive patients.These paired serum specimens were tested by serum dilution neutralization test with XJ0625 virus.Most of the samples contained neutralizing antibody with a titer of 40 or 80.The results indicated that human infection with TAHV is common in the area.Furthermore,the result of IFA test coordinate with the NT test,so it may be used as a preliminary screening test.
Sera samples of acute and convalescent phase of unknown encephalitis patients were screened by IFA test for several kinds of arbovirus.The results showed all the sera samples have IgG and IgM against flavivirus with a high titer and no positive results for non-flavirus.Combination with the features of clinical manifestations,prognostic and immunological responsds,it was concluded that pathogen of the epidemic may be a kind of unknown flavivirus.
In the study,an pathogen investigation on an encephalitis outbreak in south Xinjiang in the summer of 2004 was conducted.The results showed only 2cases had 4 fold rises among all six pairs of acute and convalescent serum specimens collected during the disease.And the neutralizing antibody titer against JEV were very low.The remaining four pairs of specimens of JEV neutralizing antibodies were negative, according to the results,it was not sufficient to take JEV as the pathogen for the endemic.So,a wide range of arbovirus antibody screening was carried out.The results showed that IgM and IgG antibody against JEV,DENV,WNV,SLEV and other flavirus were positive and negative for TAHV and other alphavirus.Taking into account serological cross-reaction between flavivirus,these results prompted only there may be some flavivirus infection.But the real pathogen is still not certain.It is recommended that NT test against a variety of flavivirus should be carried out.
In addition,641 cases of patients with unknown fever in northern and southern Xinjiang were collected in 2008 to carried out the IgM antibody screening against WNV,DENV and JEV.The results showed that 11 cases has IgM antibody against WNV,19 cases against DENV and 4 cases against JEV.These results indicated the existence of a wide range of the flavivirus in Xinjiang and may cause human diseases in summer.
The importance of the study
In summary,arbovirus investigation in Xinjang was carried out in this study during 2006,2007 and 2008.3 strains of TAHV and 41 strains of LNV were isolated from mosquitoe pools.The isolation of TAHV from southern region of Xinjiang indicated this virus prevelant in local area and no isolation from northern region of Xinjiang indicated that this virus may not active there.Molecular character of all the viruses indicated that there may be several subtypes existed in Xinjiang. Seroepidemiological results showed TAHV may be prevelant in local area and unknown fever patients may be infected by such virus.
LNV existed in most region of Xinjiang,the sequence of all the strains were relatively conserved.Phylogenetic results showed four subtypes can be classified based on segment 10.Isolates from the Xinjiang region,and Jilin exist significant regional comparison.Isolates from Aletai showed relatively high homology and the characteristic consensus sequences with Jilin isolates.It suggested that there may be similar virus between Jilin and Xinjiang.
These findings give us a clue to strengthen arbovirus surveillance in order to prevent arbovirus diseases.Identification and molecular analysis of TAHV and LNV isolates isolated from Xinjiang can help to know more about the virus characterization, especially the molecular difference from foreign isolates,and can provide basic information for further study of the arboviruses distributed in Chinese territory.
引文
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