浙江省2008-2009年三起诺如病毒胃肠炎暴发的病原分子特征分析

Objective To study the molecular characteristic of norovirus in 3 outbreaks of gastroenteritis in Zhejiang province. Methods During January 2008 and December 2009, fecal specimens of patients were collected from 3 outbreaks of acute viral gastroenteritis. Noroviruses were detected by Real-time RT-PCR. Part of the positive samples were randomly selected and detected by RT-PCR. PCR products were sequenced. Sequence analysis was undertaken based on partial sequence of RNA dependent RNA polymerase(RdRp)and capsid protein gene. Some positive samples were amplified by 3' RACE(rapid amplification of cDNA 3' ends), 3200 bp in length. The exact whole ORF2, ORF3 and 3' untranslation regions(UTR)gene of norovims were identified. Results There were in total 3 outbreaks of viral gastroenteritis caused by norovirus being reported. A total of 62 stools were obtained from cases with acute gastroentefitis. Noroviruses were detected in 41 cases including 27 strains of genogroup Ⅰ norovirus and 9 strains of genogroup Ⅱ norovirus, 5 strains of genogroup Ⅰ + Ⅱ norovirus. Four genotypes including G Ⅰ .8, G Ⅱ .b, G Ⅰ .2/0 Ⅰ .6 recombination together with co-infection of G Ⅰ .8 and G Ⅱ .b were detected. Conclusion Norovirus was confirmed as the major cause of outbreaks of viral gastroenteritis in Zhejiang province and multiple genotype of norovirus were identified from the outbreaks. It was the first time to have found a recombinant of G Ⅰ .6 capsid and G Ⅰ .2 polymerase norovims as well as the co-infection of G Ⅰ .8 and G Ⅱ .b norovirus in the same sample.     

浙江省2008-2009年三起诺如病毒胃肠炎暴发的病原分子特征分析

Objective To study the molecular characteristic of norovirus in 3 outbreaks of gastroenteritis in Zhejiang province. Methods During January 2008 and December 2009, fecal specimens of patients were collected from 3 outbreaks of acute viral gastroenteritis. Noroviruses were detected by Real-time RT-PCR. Part of the positive samples were randomly selected and detected by RT-PCR. PCR products were sequenced. Sequence analysis was undertaken based on partial sequence of RNA dependent RNA polymerase(RdRp)and capsid protein gene. Some positive samples were amplified by 3' RACE(rapid amplification of cDNA 3' ends), 3200 bp in length. The exact whole ORF2, ORF3 and 3' untranslation regions(UTR)gene of norovims were identified. Results There were in total 3 outbreaks of viral gastroenteritis caused by norovirus being reported. A total of 62 stools were obtained from cases with acute gastroentefitis. Noroviruses were detected in 41 cases including 27 strains of genogroup Ⅰ norovirus and 9 strains of genogroup Ⅱ norovirus, 5 strains of genogroup Ⅰ + Ⅱ norovirus. Four genotypes including G Ⅰ .8, G Ⅱ .b, G Ⅰ .2/0 Ⅰ .6 recombination together with co-infection of G Ⅰ .8 and G Ⅱ .b were detected. Conclusion Norovirus was confirmed as the major cause of outbreaks of viral gastroenteritis in Zhejiang province and multiple genotype of norovirus were identified from the outbreaks. It was the first time to have found a recombinant of G Ⅰ .6 capsid and G Ⅰ .2 polymerase norovims as well as the co-infection of G Ⅰ .8 and G Ⅱ .b norovirus in the same sample.     

艾滋病患者马尔尼菲青霉菌优势株酵母相全长cDNA文库的构建和初步分析

Objective To construct a full length cDNA library of the dominant strain of Penicillium marneffei (PM) in yeast phase isolated from AIDS patients in Guangdong province and screen UniGenes as well as full-length genes, so as to establish the foundation for the study of PM's functional genes and pathogenic mechanisms. Methods CloneMiner cDNA construction kit was utilized to extract mRNA of the dominant PM strain isolated from AIDS patients in Guangdong province. The mRNA was reversed into cDNA, then cloned into a pDONR222 vector by BP recombination to obtain an Uncut cDNA library, which was homogenized later to construct a normalized cDNA library with the principal of saturation hybridization for DNA genome. 2000 clones were chosen randomly to make a bi-directional sequencing and analyzed with bioinformatics for screening UniGenes and full-length genes. Results The total clone number of the Uncut cDNA library was 1.16 × 107 cfu/mL, with a recombination rate of 95% and an average insertion element being over 1 kb. The total clone number of the normalized cDNA library was 1.18 × 106 cfu/mL, with a recombination rate of 95% and an average insertion element being over 1 kb as well. 1945 genes which DNA length were longer than 1 kb were obtained by sequencing and merged into 1360 UniG enes, of which 632 genes were full-length ones. Conclusions The full-length cDNA library of the dominant strain of PM from AIDS patients in Guangdong province possesses good quality.Meanwhile, the technical routine presents high efficiency in obtaining full-length genes and establishing a gene expression spectrum, which can contentedly meet the needs of future experiments.     

浙江省1999--2010年乙型流感病毒血凝素 和神经氨酸酶基因变化特征分析

目的 分析1999-2010年浙江省乙型流感病毒主要抗原基因血凝素(HA)和神经氨酸酶(NA)的分子变异特征.方法 采集浙江省流感暴发疫情和哨点监测医院的流感样患者呼吸道标本,荧光定量RT-PCR快速检测和病毒分离,选取乙型流感病毒分离代表株进行HA和NA基因测序,采用生物信息学软件分析变异和进化.结果 共分析浙江省乙型流感病毒分离株34株,其中Victoria系20株,Yamagata系14株;1999-2010年间Victoria系毒株的HA1基因变异率4.5%,Yamagata系毒株为3.4%;2004年后分离的Victoria系毒株均为基因重配株,HA属于Victoria 系,而NA属于Yamagata系;2010年新型甲型HINI流感流行高峰过后,浙江省仍以乙型流感病毒流行为主,分离株与2009-2010年流感疫苗株B/Brisbane/60/2008接近,与往年乙型流感毒株相比HA和NA发生多个氨基酸位点变异.结论 1999-2010年浙江省乙型流感病毒流行株发生明显变异,基因重配和抗原漂移是病毒发生变异的主要机制.

Abstract：

Objective To Characterize the genetic diversity of hemagglutinin(HA)and neuraminidase(NA)of influenza B viruses isolated in Zhejiang province during 1999-2010.Methods Respiratory specimens were collected from patients with flu-like syndrome during the influenza outbreaks or from the hospitals which carrying out influenza surveillance project in Zhejiang province.Samples were detected by real-time RT-PCR and isolated for influenza virus.HA1 and NA genes of influenza B virus isolates were amplified and sequenced.Phylogenetic comparison and genetic diversity analysis were performed using the bioinformation software.Results A total of 34 influenza B viruses were evolved in this study including Victoria-like and Yamagata-like strains according to the results of the HI test.The mutation rate of Victoria-like HA1 gene was 4.5% and Yamagata-like HA1 gene was 3.4%,respectively.The Victoria-like influenza B isolates had appeared to be all re-assortants having a Victoria lincage HA and Yamagata lineage NA since 2004.The predominant type of influenza virus isolates in 2010 was also influenza B virus after the H1N1 flu pandemic in Zhejiang province.The isolated strains were antigenicaily and genetically similar to B/Brisbane/60/2008--the vaccine strain proposed for 2009-2010.Many difierences of HA1 and NA amino acids existed in the current isolates when compared to previous influenza B strains.Conclusion Significant diversity was generated among influcnza B virus isolated from 1999 to 2010 in Zhejiang province.Genetic re-assortment and antigenic drift seemed the main evolutionary mechanism on influenza B virus.     

浙江省嘉兴地区初孕妇女自然流产状况研究

Objective To study the current status of spontaneous abortion of primigravid women in Jiaxing areas of Zhejiang province of China. Methods We analyzed the data from both perinatal healthcare surveillance program and spontaneous abortion, collected in Jiaxing areas by the Institute of Reproductive and Child Health, Peking University. The study population consisted of 14 769 primigravid women (excluding induced abortion, ectopic pregnancy and molar pregnancy as outcomes) attempting to become pregnant who registered between 1993 and 1995. Results 1454 spontaneous abortion cases were identified, with the spontaneous abortion rate as 9.8% (95% CI:9.3%-10.3%). The mean gestational weeks at pregnancy diagnosis were 7.6±2.1 weeks, the mean gestational weeks at miscarriage were (10.1±3.1) weeks and the incidence of first-trimester (≤12 weeks) spontaneous abortion was 7.3%(95%CI: 6.8%-7.7%), accounting for 73.7% of all the spontaneous abortion cases. A peak for risk of miscarriage was around 8-13 weeks, accounting for 37.7% of all spontaneous abortion. The observed multiple Cox regression model showed that increased spontaneous abortion rates were observed in women with age at pregnancy ≥30, being peasants and with higher education level. Conclusion The spontaneous abortion rate of primigravid women in Jiaxing areas was higher than in other areas of China. The maximum occurrence of spontaneous abortions was during period of 8-13 gestation weeks.     

浙江省嘉兴地区初孕妇女自然流产状况研究

Objective To study the current status of spontaneous abortion of primigravid women in Jiaxing areas of Zhejiang province of China. Methods We analyzed the data from both perinatal healthcare surveillance program and spontaneous abortion, collected in Jiaxing areas by the Institute of Reproductive and Child Health, Peking University. The study population consisted of 14 769 primigravid women (excluding induced abortion, ectopic pregnancy and molar pregnancy as outcomes) attempting to become pregnant who registered between 1993 and 1995. Results 1454 spontaneous abortion cases were identified, with the spontaneous abortion rate as 9.8% (95% CI:9.3%-10.3%). The mean gestational weeks at pregnancy diagnosis were 7.6±2.1 weeks, the mean gestational weeks at miscarriage were (10.1±3.1) weeks and the incidence of first-trimester (≤12 weeks) spontaneous abortion was 7.3%(95%CI: 6.8%-7.7%), accounting for 73.7% of all the spontaneous abortion cases. A peak for risk of miscarriage was around 8-13 weeks, accounting for 37.7% of all spontaneous abortion. The observed multiple Cox regression model showed that increased spontaneous abortion rates were observed in women with age at pregnancy ≥30, being peasants and with higher education level. Conclusion The spontaneous abortion rate of primigravid women in Jiaxing areas was higher than in other areas of China. The maximum occurrence of spontaneous abortions was during period of 8-13 gestation weeks.     

,China)[Objective] To understand the spe

ricles were scraped off by a bistoury at first and then pre-served in 70% of ethanol.Hoyer's medium was used to mount the chigger mites onto glass slides and the specimens of the chigger mites on the slides were finally identified into species under a microscope.Dispersion coefficient,clumping index,cassie index and patchiness index were used to analyze the spatial patterns of Chigger Mites.[Results] The captured 302 small mammals were identified as 16 species and 13 genera in 7 families.4 550 individuals of Chigger Mites were collected from the two auricles of 302 small mammal hosts and then they were identified as 63 species and 12 genera in     

浙江省2008-2009年三起诺如病毒胃肠炎暴发的病原分子特征分析

目的 分析浙江省3起病毒性胃肠炎暴发疫情的诺如病毒分子特征.方法 收集监测期间病毒性胃肠炎暴发疫情患者的粪便标本,采用荧光定量RT-PCR方法检测诺如病毒,并选择部分阳性标本扩增部分多聚酶区(RdRp)和衣壳蛋白区,同时采用3′RACE(rapidamplification of cDNA 3′ends)扩增诺如病毒基因组3′末端,获得完整的开放读码框架(ORF)2和ORF3序列.结果 3起暴发疫情共检测标本62份,诺如病毒阳性41份,其中诺如病毒Ⅰ(G Ⅰ)基因组阳性27例,Ⅱ基因组(GⅡ)阳性9例,G Ⅰ+GⅡ阳性5例.结果 显示,引起浙江省2008-2009年3起病毒性胃肠炎暴发疫情的诺如病毒具有病毒基因型的多样性,包括G Ⅰ.8、GⅡ.b、GⅠ.2与GⅠ.6重组株、GⅠ.8和GⅡ-b混合感染.结论 诺如病毒是浙江省病毒性腹泻暴发疫情的重要病原体,呈现出病毒基因型的多样性,并在省内首次检测到诺如病毒的重组和混合感染毒株.

Abstract：

Objective To study the molecular characteristic of norovirus in 3 outbreaks of gastroenteritis in Zhejiang province. Methods During January 2008 and December 2009, fecal specimens of patients were collected from 3 outbreaks of acute viral gastroenteritis. Noroviruses were detected by Real-time RT-PCR. Part of the positive samples were randomly selected and detected by RT-PCR. PCR products were sequenced. Sequence analysis was undertaken based on partial sequence of RNA dependent RNA polymerase(RdRp)and capsid protein gene. Some positive samples were amplified by 3' RACE(rapid amplification of cDNA 3' ends), 3200 bp in length. The exact whole ORF2, ORF3 and 3' untranslation regions(UTR)gene of norovims were identified. Results There were in total 3 outbreaks of viral gastroenteritis caused by norovirus being reported. A total of 62 stools were obtained from cases with acute gastroentefitis. Noroviruses were detected in 41 cases including 27 strains of genogroup Ⅰ norovirus and 9 strains of genogroup Ⅱ norovirus, 5 strains of genogroup Ⅰ + Ⅱ norovirus. Four genotypes including G Ⅰ .8, G Ⅱ .b, G Ⅰ .2/0 Ⅰ .6 recombination together with co-infection of G Ⅰ .8 and G Ⅱ .b were detected. Conclusion Norovirus was confirmed as the major cause of outbreaks of viral gastroenteritis in Zhejiang province and multiple genotype of norovirus were identified from the outbreaks. It was the first time to have found a recombinant of G Ⅰ .6 capsid and G Ⅰ .2 polymerase norovims as well as the co-infection of G Ⅰ .8 and G Ⅱ .b norovirus in the same sample.     

云南省伤寒副伤寒空间分布特征及其气候影响因素研究

Objective To characterize the spatial distribution of typhoid and paratyphoid fever(TPF)in Yunnan province, China and to determine the effectiveness of meteorological factors on the epidemics of TPE Methods Data of reported TPF cases in Yunnan province(2001 -2007)from the China Information System for Diseases Control and Prevention was applied to GIS-based spatial analyses to detect their spatial distribution and clustering of TPF incidence at the county level.Panel data analysis was used to identify the relationships between the TPF incidence and meteorological factors including monthly average temperature, monthly cumulative precipitation and monthly average relative humidity. Results During the study period, the average incidence of TPF in Yunnan province was 23.11/100 000, with majority of the TPF cases emerged in summer and autumn. Although widely distributed, two TPF clusters were detected in Yunnan province based on the spatial analysis:one area around Yuxi city with the average annual incidence as 207.45/100 000 and another at the junctions of Yunnan province with Burma and Laos. Based on results from panel data analysis, the incidence of TFP was shown to be associated with meteorological factors such as temperature,precipitation, relative humidity and one month lag of temperature increase [10 ℃ increase in the monthly average temperature:IRR=1.30(95%CI: 1.24-1.36);10% increase in monthly average relative humidity:IRR= 1.07(95%CI: 1.05-1.09); 100 mm rise in monthly cumulative precipitation:IRR=1.02(95%CI: 1.00-1.03); and 10 ℃ average temperature increase, the last month: IRR=1.73(95%CI: 1.64-1.82)]. Conclusion Areas with high TPF incidence were detected in this study,which indicated the key areas for TPF control in Yunnan province. Meteorological factors such as temperature, precipitation and humidity played a role in the incidence of TPF.     

云南省伤寒副伤寒空间分布特征及其气候影响因素研究

Objective To characterize the spatial distribution of typhoid and paratyphoid fever(TPF)in Yunnan province, China and to determine the effectiveness of meteorological factors on the epidemics of TPE Methods Data of reported TPF cases in Yunnan province(2001 -2007)from the China Information System for Diseases Control and Prevention was applied to GIS-based spatial analyses to detect their spatial distribution and clustering of TPF incidence at the county level.Panel data analysis was used to identify the relationships between the TPF incidence and meteorological factors including monthly average temperature, monthly cumulative precipitation and monthly average relative humidity. Results During the study period, the average incidence of TPF in Yunnan province was 23.11/100 000, with majority of the TPF cases emerged in summer and autumn. Although widely distributed, two TPF clusters were detected in Yunnan province based on the spatial analysis:one area around Yuxi city with the average annual incidence as 207.45/100 000 and another at the junctions of Yunnan province with Burma and Laos. Based on results from panel data analysis, the incidence of TFP was shown to be associated with meteorological factors such as temperature,precipitation, relative humidity and one month lag of temperature increase [10 ℃ increase in the monthly average temperature:IRR=1.30(95%CI: 1.24-1.36);10% increase in monthly average relative humidity:IRR= 1.07(95%CI: 1.05-1.09); 100 mm rise in monthly cumulative precipitation:IRR=1.02(95%CI: 1.00-1.03); and 10 ℃ average temperature increase, the last month: IRR=1.73(95%CI: 1.64-1.82)]. Conclusion Areas with high TPF incidence were detected in this study,which indicated the key areas for TPF control in Yunnan province. Meteorological factors such as temperature, precipitation and humidity played a role in the incidence of TPF.     

浙江省天台县2011－2018年汉坦病毒基因型别和进化变异研究

目的 研究2011－2018年肾综合征出血热（HFRS）国家监测点浙江省天台县汉坦病毒（HV）的基因型别和进化变异情况，了解天台县HV分子流行病学特点。方法 将天台县2011－2018年HV抗原阳性的鼠肺标本超声后提取核酸，利用HV型特异性引物，应用巢式PCR对部分M片段进行扩增分型和序列测定，将天台县2011－2018年的HV序列与国内外其他已知的HV序列进行比较，以明确该地区的基因型别，分析病毒的进化变异情况。结果 HV抗原阳性的67份鼠肺标本经型特异性引物巢式PCR扩增后31份标本为阳性，其中30份为汉滩病毒（HTNV）、1份为汉城病毒（SEOV），31份PCR阳性鼠肺均来自黑线姬鼠。31份巢式PCR阳性产物的部分M片段核苷酸序列有30株分布在HTNV单元群，1株分布在SEOV单元群。HTNV单元群中的天台T2018-130株与天台其余29株及国内外其他株同源性为84.8%～87.9%，差异较大，天台其余29株亲缘关系较近； SEOV发生群中的T2016-31株来自黑线姬鼠的鼠肺标本，与SEOV天台以往分离株ZT71株、ZT10株和浙江温州分离株Z37株在系统发生树上分布于同一或临近分支。结论 浙江省天台县HV流行的主要型别HTNV基因表现出明显的地理聚集现象，但也存在着基因差异较大的变异株T2018-130株；同时从病毒基因序列分析证实SEOV T2016-31株存在于黑线姬鼠中，可能意味着在SEOV进化过程中病毒在宿主间发生了“溢出”现象。     

浙江省丽水地区2株汉坦病毒分离株的型别鉴定和基因差异研究

目的 从浙江省丽水地区肾综合征出血热(HFRS)疫区阳性鼠肺标本中分离汉坦病毒(HV),并对丽水分离株进行分子生物学鉴定,分析M和S片段基因序列以确定毒株的型别和基因差异程度.方法 收集宿主动物标本,采用直接免疫荧光检测鼠肺标本HV抗原.将HV抗原阳性的鼠肺标本接种Vero-E6细胞,分离HV.提取病毒总RNA,应用反转录聚合酶链反应(RT-PCR)扩增病毒M和S基因全片段,克隆入质粒载体,纯化后测定序列,并同HV其他病毒株进行比较.结果 成功分离到2株HV,扩增出2株病毒的M和S全片段并测定了序列,经对核苷酸序列分析表明,2株病毒与现有的HTNV有最高的同源性,均为HTNV,但与国内外其他的HTNV核苷酸差异高达13.4%～20.7%和10.3%～16.1%.用M和S片段核苷酸序列所构建的系统进化树显示,2毒株分在HTNV发生群,与HTNV的Z10、A9、Z5病毒株亲缘关系最近,但独自构成一进化支.结论 浙江丽水分离株可以定型为HTN型病毒,与国内外其他的HTN型病毒基因差异较大.     

湖南省啮齿动物携带汉坦病毒的分子流行病学研究

目的研究湖南省啮齿动物中汉坦病毒(HV)流行情况及病毒型别。方法采用IFA法检测鼠肺中HV抗原;用RT-PCR法扩增阳性标本中HV的部分S和M片段;构建系统发生树进行系统发生分析及分型。结果在湖南省HV流行的地区共捕获啮齿动物344只,其中6份标本HV抗原阳性,病毒携带率为1.74%。对扩增出的部分S与M片段的核苷酸序列分析表明,5份标本中的病毒为汉城型HV(SEOV),1份为汉滩型HV(HTNV)。用S(620～990 nt)与M片段G2区(2001～2301 nt)核苷酸序列所构建的系统进化树显示,湘乡褐家鼠、黄胸鼠及黄毛鼠携带的病毒均为SEOV的S4亚型;宁远褐家鼠携带的病毒为SEOV的一个新亚型;石门小家鼠携带的病毒为HTNV的H4亚型。结论湖南省为混合型HV疫区,以汉城病毒为主,并具有宿主多样性。     

2012 - 2015年江西省啮齿动物间流行的汉坦病毒基因型和基因亚型分析

目的 了解2012 - 2015年江西省啮齿动物间流行的汉坦病毒基因型别和基因亚型。方法 选取江西省肾综合征出血热疫情发生地区捕鼠,收集鼠肺标本,采用直接免疫荧光法检测鼠肺中汉坦病毒抗原,提取阳性鼠肺标本核酸,设计并采用特异性引物分别对病毒的S、M、L 3个基因进行RT - PCR扩增,对扩增产物测序和分析。结果 共捕鼠2 235只,其中汉坦病毒阳性的86只,阳性率3.85%,流行的HV基因型为汉滩型(Hantaan orthohantavirus,HTNV)和汉城型（Seoul orthohantavirus,SEOV）。江西省HTNV之间S、M、L 3个基因核苷酸的同源性为93.4%～100%,与国内外HTNV参考株3个基因核苷酸的同源性为77.3%～88.0%,在3个基因系统进化树上均呈独立分支;M基因系统进化分析发现江西省SEOV分布在S3、S4亚型分支以及1个独立的进化分支,该独立分支的9株SEOV与国内外SEOV M基因核苷酸同源性仅为84.3%～88.7%。结论 江西省啮齿动物流行的HV为HTNV和SEOV 2种基因型别,其中HTNV为新的基因亚型,SEOV存在3个基因亚型:S3、S4和1个新的基因亚型。     

浙江省温州市啮齿动物中汉坦病毒的分子流行病学研究

目的 研究浙江省温州市啮齿动物中汉坦病毒(HV)流行情况及病毒型别,为该地区肾综合征出血热(HFRS)的预防控制提供科学依据.方法 采用间接免疫荧光法(IFA)检测鼠肺中HV抗原;用RT-PCR法扩增阳性样品中HV的部分S片段及部分M片段;构建系统发生树进行系统发生分析及分型.结果在温州市HFRS疫区共捕获啮齿动物96只,在6份鼠肺样品中检测到HV抗原,其中4只褐家鼠,1只黄胸鼠与1只黄毛鼠,病毒携带率为6.3%.用汉城病毒(SEOV)特异引物从其中5份HV抗原阳性样品中扩增出部分S片段(620～999nt)及部分M片段(2001～2301nt)并测定序列.对扩增出的部分S及M片段的核苷酸序列分析发现,5株病毒与现有的SEOV有高同源性,均为汉城型HV.但在用部分S片段及部分M片段核苷酸序列所构建的系统进化树上,5株病毒的聚集模式不同.结论温州市的褐家鼠、黄胸鼠、黄毛鼠均携带汉城型HV,并可能发生基因片段的重排.     

2007年浙江省鼠类感染汉坦病毒监测及病毒分离

目的了解浙江省鼠类自然感染汉坦病毒（HV）情况和病毒型别,为肾综合征出血热（HFRS）防治提供科学依据。方法采集鼠类肺组织和血清标本,用间接免疫荧光试验检测鼠血清IgG抗体,直接免疫荧光试验检测鼠肺Hv抗原。HV抗原阳性鼠肺接种Vero—E6细胞分离病毒,用HV单克隆荧光抗体鉴定分离株的型别。结果共捕获鼠形动物1129只,其中黑线姬鼠为优势种,鼠肺HV抗原阳性率为3．0％,鼠血标本IgG抗体阳性57份,阳性率8．0％。分离到6株汉滩（HTN）型病毒,其中来自黑线姬鼠5株,褐家鼠1株。结论浙江省存在以黑线姬鼠为主要传染源的HFRS疫源地,HV主要流行型别为HTN型。     

北京市城区鼠间汉坦病毒感染的流行病学研究

目的了解北京城区中心地带鼠间汉坦病毒（Hv）感染状况及其病毒型别差异。方法选取北京城区某方圆约1km两街区之内的食物资源丰富、场所隐蔽的中、小餐饮行业、蔬菜集贸市场及周边垃圾堆、卫生死角等生态环境，从2003年12月至2004年11月每月连续采用夹夜法捕鼠，计算鼠密度和鼠种构成。应用间接ELISA法检测鼠血清IgG抗体；针对汉坦病毒M基因部分片段设计汉城型汉坦病毒（SEOV）特异性引物，应用RT-PCR法检测宿主动物携带HV情况，阳性标本直接测序。利用Excel和SPSS软件分析宿主动物HV感染的动态特征。最后用DNASTAR软件对获得序列进行比较分析。结果共捕获啮齿动物229只，平均鼠密度6．41％（229／3570），褐家鼠和小家鼠分别为餐饮业、农贸市场生境的优势鼠种。褐家鼠血清抗体阳性率3．51％（6／171），带病毒率为0．58％（1／171）；小家鼠血清抗体阳性率为6．90％（4／58），带病毒率为172％（1／58）。病毒检出率以11、12月最高，抗体水平冬春季（1、4月）最高，与春季人群肾综合征出血热（HFRS）流行季节基本一致。2份扩增阳性标本（分别来源于小家鼠和褐家鼠）均为SEOV，差异为5．40％，系统发育分析显示他们位于2个不同的较小分支，但均位于Z37支系。结论北京城区2种主要的HV宿主在一些鼠害重点行业密度仍较高，虽然HV基础感染率不高，但持续存在，为春季人群HFRS发病高峰的重要影响因素。来自2个不同宿主的HV基因有一定的差异。小家鼠的传播作用有所加强，应予重视。     

北京市部分人群和农产品集散地鼠间汉坦病毒感染的研究

目的了解北京地区鼠间汉坦病毒（HV）感染的可能来源与线索.方法2004年4-5月选取北京城区某大型火车站和郊区某大型农产品贸易集散地,采用夹夜法捕鼠,应用间接ELISA法检测鼠血清IgG抗体;针对HV M和S基因片段设计汉城型病毒（SEOV）特异引物,应用逆转录-聚合酶链反应检测鼠肺中HV,阳性标本直接测序,与GenBank已知序列进行两两比较,对获得序列进行多重比较分析并构建系统发育树.结果共捕获鼠类24只,平均密度3.49%（24/690）,火车站和农产品集散地平均鼠密度分别为1.25%（3/240）和4.67%（21/450）,血清抗体阳性率分别为0.0%和9.5%.优势种分别为小家鼠和褐家鼠,带病毒率分别为14.28%（1/7）和5.89%（1/17）.序列比较结果显示：2份扩增阳性标本M片段核苷酸差异为7.6%,来自火车站的dc501与山东省SD227、河北省Hebei4株亲缘关系较近.来自集贸市场的BjFT01株与GenBank中已注册的其他北京市SEOV株比较,存在一些独特的基因差异,并与海南省HN71株、浙江省K24-E7株多个位点变异类型一致,同源性较高,构成一个支系.而其S片段与韩国80-39株同源性最高（96.4%）,与浙江K24-E7同源性为95 5%.结论北京市人群及农产品集散地鼠类HV感染对肾综合征出血热流行的潜在危险较大,家栖鼠携带并传播的SEOV由上述类似地区从外埠输入到北京的可能性较大,但有待进一步研究.     

浙江省慈溪市啮齿动物中汉坦病毒分子流行病学研究

目的 研究浙江省慈溪市啮齿动物中汉坦病毒(HV)流行情况及病毒型别.方法 采用间接免疫荧光法(IFA)检测鼠肺中HV抗原;用逆转录聚合酶链反应(RT-PCR)法扩增阳性样品中HV的部分S片段;构建系统发生树进行系统发生分析及分型.结果 在慈溪市肾综合征出血热(HFRS)疫点共捕获啮齿动物243只,在7份鼠肺样品中检测到HV抗原,其中4只为褐家鼠,3只为黄胸鼠,病毒携带率为2.88%.用汉城型病毒(SEOV)特异性引物从6份HV抗原阳性样品中扩增出部分S片段(620～999 nt)并测定了序列.对扩增出的部分S片段核苷酸序列分析表明,6株病毒与现有的SEOV有最高的同源性,均为SEOV.用部分S片段核苷酸序列所构建的系统进化树显示,6株病毒构成两个单元支,其中由褐家鼠携带的3株病毒与BjHD01株病毒的亲缘关系最近,分在同一分支,由黄胸鼠携带的3株病毒构成另一分支,与L99、R22及Hb8610株的亲缘关系较近.结论 慈溪市的褐家鼠与黄胸鼠分别携带一种不同亚型的SEOV,表现出SEOV在同一地区的遗传多样性,并支持宿主与汉坦病毒共进化的理论.     

辽宁省汉滩型汉坦病毒基因特征及分布研究

目的探讨辽宁省汉滩型汉坦病毒(HTNV)的基因特征及其分布情况。方法在辽宁省肾综合征出血热(HFRS)主要流行地区收集鼠肺和HFRS患者标本,采用间接免疫荧光法检测鼠肺中HV抗原,以RTPCR方法扩增标本中M和S基因片段的特异性核苷酸序列,将HTNV的阳性产物测序,并进行同源性比对和系统发生分析。结果辽宁省HFRS疫区HTNV主要为黑线姬鼠所携带;从鼠肺标本中扩增出M片段4份,患者标本中扩增出M片段5份,S片段1份;M片段的核苷酸同源性分析表明,辽宁株与Bao14、CJAp267等株同源性最高,为95%～97%,与HTNV原型株76-118同源性为87.4%～89.0%;系统发生分析显示辽宁株均分布于同一支内,与Bao14、CJAp267等株构成一个独立的支系,同属于H4亚型;TL2S基因片段与YaluRiver13核苷酸同源性最高(97.1%),与HTNV原型株76-118同源性为91.2%;推导的S片段氨基酸同源性,TL2与Bao14、CJAp93同源性较高,为93.0%～96.2%,与其他代表株同源性多在74.1%～81.6%;而且基于S片段的系统发生分析提示与M片段的分型结果基本一致,与Bao14、CJAp93等株位于同一分支内,为H4亚型。结论目前辽宁省流行的HTNV主要为H4基因亚型,基因亚型的分布相对比较单一。