登革病毒2型广东省分离株的鉴定及NS1基因序列分析  

Identification and NS1 gene sequence analysis of dengue virus 2 Guangdong strains

作　　者：方美玉[1] 赵文忠[1] 刘建伟[1] 白志军[1] 田小东[1] 林立辉[1] 

机构地区：[1]广州军区军事医学研究所,510507

出　　处：《中华传染病杂志》2001年第1期23-26,共4页Chinese Journal of Infectious Diseases

基　　金：全军"九五"医学科研基金资助项目(97L059);广东省"五个一科教兴医工程"重点项目基金资助项目

摘　　要：目的　明确广东省南海市 1998年夏、秋季一批发热、出疹患者的诊断 ,并从基因水平分析其流行株的来源。方法　收集疑似登革热 (DF)患者血清 5 2份 ,应用逆转录 聚合酶链反应 (RT PCR)、病毒分离和间接免疫荧光技术分别进行了病原学和血清学检测 ;同时采用分子克隆技术将PCR扩增产物克隆到T载体 ,并进行序列测定。结果　从 2 5份发病早期患者血标本中分离病毒 10份 ,经用单克隆抗体间接免疫荧光检测和RT PCR检测证实为登革病毒 2型感染。基因序列分析表明 ,1998年广东分离的登革 2型病毒与ThNH P2 8/ 93、C0 16 6、16 6 81、NGC、JAM、0 4、GD0 6 / 93和S1株核苷酸的同源性分别是 :98%、97%、96 %、96 %、93%、92 %、93 %、91%。结论　此次南海市登革热暴发流行为登革病毒 2型感染所致。基因序列分析提示 :1998年广东省南海市分离的登革病毒Objective To identify and diagnose the patients with suspected Dengue fever in Nan Hai city, Guangdong province, in 1998, and analyze the gene sequence of Dengue virus (DEN) Guangdong strains as well as compare their homology and explore their sources. Methods Serum samples from 52 patients with suspected dengue fever were assayed by aetiologycal and serological detection. Fragments of 413 bp from NS1 gene from DEN Guangdong strains were amplified by reverse transcription and polymerase chain reaction (RT PCR), then cloned into the PGEM T vectors and sequenced. Results Ten strains of virus were isolated from 25 patients in the early stage of the disease. Dengue virus (DEN) 2 were identified by monoclonal antibody immunofluorescence assay (IFA) and reverse transcription polymerase chain reactions (RT PCR). The results of sequence analysis showed that the nucleotide sequence homologies of the strains isolated in 1998 among the ThNH P28/93, C0166, 16681, NGC, JAM, 04, GD06/93, S1 were 98%,97%,96%,96%,93%,92%,93% and 91% respectively. Conclusion It shows that the outbreak of dengue fever in Nan Hai city of Guangdong province was caused by DEN 2 type. The result of DNA sequence analysis suggests that the Guangdong DEN 2 local strains isolated in 1998 were similar to those from Thailand.

关 键 词：登革热病毒 逆转录-聚合酶链反应 序列分析 DNA 序列同源性 

分 类 号：R373.33[医药卫生—病原生物学]