中华流行病学杂志
中華流行病學雜誌
중화류행병학잡지
CHINESE JOURNAL OF EPIDEMIOLOGY
2009年
2期
175-178
,共4页
姚苹苹%朱函坪%徐芳%王志刚%梅玲玲%雷永良%谢荣辉%王复甦%朱智勇%邓小昭%张云
姚蘋蘋%硃函坪%徐芳%王誌剛%梅玲玲%雷永良%謝榮輝%王複甦%硃智勇%鄧小昭%張雲
요평평%주함평%서방%왕지강%매령령%뢰영량%사영휘%왕복소%주지용%산소소%장운
肾综合征出血热%汉坦病毒%基因型%系统发生分析
腎綜閤徵齣血熱%漢坦病毒%基因型%繫統髮生分析
신종합정출혈열%한탄병독%기인형%계통발생분석
Hemorrhagic fever with renal syndrome%Hantavirus%Genotype%Phylogenetic analysis
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.
目的 從浙江省麗水地區腎綜閤徵齣血熱(HFRS)疫區暘性鼠肺標本中分離漢坦病毒(HV),併對麗水分離株進行分子生物學鑒定,分析M和S片段基因序列以確定毒株的型彆和基因差異程度.方法 收集宿主動物標本,採用直接免疫熒光檢測鼠肺標本HV抗原.將HV抗原暘性的鼠肺標本接種Vero-E6細胞,分離HV.提取病毒總RNA,應用反轉錄聚閤酶鏈反應(RT-PCR)擴增病毒M和S基因全片段,剋隆入質粒載體,純化後測定序列,併同HV其他病毒株進行比較.結果 成功分離到2株HV,擴增齣2株病毒的M和S全片段併測定瞭序列,經對覈苷痠序列分析錶明,2株病毒與現有的HTNV有最高的同源性,均為HTNV,但與國內外其他的HTNV覈苷痠差異高達13.4%～20.7%和10.3%～16.1%.用M和S片段覈苷痠序列所構建的繫統進化樹顯示,2毒株分在HTNV髮生群,與HTNV的Z10、A9、Z5病毒株親緣關繫最近,但獨自構成一進化支.結論 浙江麗水分離株可以定型為HTN型病毒,與國內外其他的HTN型病毒基因差異較大.
목적 종절강성려수지구신종합정출혈열(HFRS)역구양성서폐표본중분리한탄병독(HV),병대려수분리주진행분자생물학감정,분석M화S편단기인서렬이학정독주적형별화기인차이정도.방법 수집숙주동물표본,채용직접면역형광검측서폐표본HV항원.장HV항원양성적서폐표본접충Vero-E6세포,분리HV.제취병독총RNA,응용반전록취합매련반응(RT-PCR)확증병독M화S기인전편단,극륭입질립재체,순화후측정서렬,병동HV기타병독주진행비교.결과 성공분리도2주HV,확증출2주병독적M화S전편단병측정료서렬,경대핵감산서렬분석표명,2주병독여현유적HTNV유최고적동원성,균위HTNV,단여국내외기타적HTNV핵감산차이고체13.4%～20.7%화10.3%～16.1%.용M화S편단핵감산서렬소구건적계통진화수현시,2독주분재HTNV발생군,여HTNV적Z10、A9、Z5병독주친연관계최근,단독자구성일진화지.결론 절강려수분리주가이정형위HTN형병독,여국내외기타적HTN형병독기인차이교대.
Objective To isolate hantavirus from Lishui county-one of the epidemic regions for hemorrhagic fever with renal syndrome(HFRS),in Zhejiang province,and to identify the serotype and molecular/biological characteristics of a new HTN subtype hantavirus(HV)strains,hopefully to provide evidence for HFRS prevention and therapy.Methods Data on the host animals was collected from Lishui,Zhejiang province in 2007.Direct immunofluorscece assay was adopted to determine HFRS antigens and the lung tissues from HV infeeted Vero-E6 cells for HV isolation,then total RNA was extracted from Hantavirus Lishui strains and amplified by RT-PCR M,S segments of strains genome were also clened and sequenced and compared with those of other strains of HV Results 2 strains virus(ZLS6-11 and ZLS-12)Were successfully isolated from 7 positive lung samples of mice and were identified as HTNV by anti-McAb and phylogenetic analysis.With sequence compation.We found that 2 strains with complete M and S segment had higher homology with HTN-type strains than with other types of HV,but 13.4%-20.7%and 10.3%-16.1%of the genes were found which were difierent from HTNV.The phylogenetic trees constructed by complete S and M segment showed that ZLS6-11 and ZLS-12 strains were located in HTNV group,and structured independent embranchment.Conclusion ZLS6-11 and ZLS-12 Strains were believed to belong to HTN-type and phylogenetically different from the HTNV.
