广东省江门地区登革病毒分离株的鉴定及结构蛋白序列分析  

Isolation, identification and sequence analyses of dengue virus type 2 strain GD19/2001

作　　者：任瑞文[1] 方美玉[1] 洪文燕[1] 黄宝明[2] 蒋廉华[1] 刘建伟[1] 田小东[1] 程刚锋[1] 

机构地区：[1]广州军区联勤部军事医学研究所微生物研究室,510507 [2]广东省江门市卫生防疫站

出　　处：《中华流行病学杂志》2003年第4期288-290,共3页Chinese Journal of Epidemiology

基　　金：全军医学科研"十五"计划重大课题资助项目( 0 1Z0 14 )

摘　　要：目的　对广东省江门地区 2 0 0 1年夏、秋季一批发热、出疹患者进行确诊 ,并从分子水平分析流行株的可能来源。方法　分别采用免疫荧光、细胞毒力、乳鼠毒力实验以及逆转录 聚合酶链反应 (RT PCR)技术进行病毒鉴定 ,并对其结构蛋白基因序列进行克隆、测序及同源性搜索。结果37份患者血清登革病毒 (DV)IgM抗体阳性率为 97% (36 37) ,IgG抗体阳性率为 59% (2 2 37) ,最高抗体滴度均可达 1∶640。所得病毒可致C6 36细胞病变 ,具有乳鼠神经毒力 ;其结构基因序列长度为2 32 5bp ,编码 774个氨基酸 ;与其他登革 2型病毒株TSV0 1、GD0 6 93、NGC、44、ThNH、0 4、GD0 8 98及S1进行比较 ,其核酸序列同源性 (% )分别为 98、96、94、94、92、92、92、91。结论　 2 0 0 1年江门地区登革热流行为登革 2型病毒感染所致 。Objective To identify the virus isolated from Jiangmen, Guangdong province and to discuss the possible origin. Methods Using characteristics of indirect fluorescent antibody tests (IFA), reverse transcription polymerase chain reaction (RT PCR), mouse neurovirulence and cell culture to identify the isolated virus. According to the nature of dengue virus type 2 NGC strain, two pairs of primers were designed. The structural protein gene of isolated dengue virus type 2 strain was then amplified by RT PCR, cloned into pMD18 T vector and sequenced. Results Tweenty two of 37 serum samples showed a positive reaction to dengue antibody IgG, and 36 of 37 with IgM with the highest antibody titer 1∶640 . Ten samples were resulted in a cytopathy on C6/36 cells and showed a neurovirulence in suckling mice when inoculated intracerebrally. The structural gene of new isolate GD19/2001 containing 2 325 nucleotides which encoded 774 amino acids. Data on nucleotide homology were 98%, 96%, 94%,94%,92%,92%,92% and 91% compared with TSV01, GD06/93, NGC and 44, ThNH81/93, 04 and GD08/98, and S1 respeectively. Conclusion The isolated virus from Jiangmen, Guangdong province belonged to dengue virus type 2, which might come from Australia.

关 键 词：登革病毒 病毒分离 序列分析 

分 类 号：R373[医药卫生—病原生物学]