珠海登革病毒分离株的型别鉴定与序列分析研究  

Serotype Identification and Phylogenetic Sequence Analysis on Dengue Virus in Zhuhai

作　　者：马洪波[1] 朱汉荣[1] 孙虹[1] 杜坚[1] 谭华[1] 杨泽[1] 林继灿[1] 陈竞帆[2] 

机构地区：[1]珠海出入境检验检疫局,珠海519020 [2]华南农业大学生命科学学院,广州510642

出　　处：《中国国境卫生检疫杂志》2006年第B08期51-54,共4页Chinese Journal of Frontier Health and Quarantine

基　　金：珠海市科技计划项目(PC20041058)

摘　　要：[目的]了解珠海市部分人群和入境船员中登革热的流行情况和珠海市流行的登革病毒株的型别和序列特征。[方法]分别采用胶体金快速法、酶联免疫吸附试验(ELISA)对登革热抗体进行检测；采用细胞培养和逆转录一聚合酶链反应(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、测序及同源性分析。[结果]来自东南亚国家船舶的船员血清中登革病毒IgM抗体检出率为4．40％,IgG抗体为20．33％,其阳性样本经RT-PCR扩增,结果为阴性。而对本室保存的2001年珠海散发的登革热患者阳性血清,经用C6/36细胞培养分离出1株登革病毒,用登革病毒通用引物和Ⅱ型特异性引物,扩增出相应的片段,证实为登革Ⅱ型病毒；该分离株(DV2-1)的基因序列与其他9株登革Ⅱ型病毒进行系统发育关系的分析,结果显示分离株(DV2-1)与GD08/98株、FJ11/99株、16681株位于相近的分支,其中与1998年广东流行株GD08/98株系统进化关系最近。[结论]2001年珠海地区登革热的散发流行为登革Ⅱ型病毒所致,推测其可能是输入性传染。Objective To analyze the epidemic characters of Dengue fever in certain group of peoples in Zhuhai city and some entry crew from south - east countries. Both the samples collected for identifying the serotype and phylogenetis characteristic of Dengue virus. Methods IgG/IgM antibodies to the Dengue virus were detected by InstaTestTM Dengue and ELISA. Virus isolation and identification were performed by cell culture and further using RT - PCR to identify the serotype of Dengue Virus. The PCR product was cloned to pGEM - T Easy, and sequenced and blasted in the Genbank to identify the isolated strain. The phylogenetic tree was produced by using Clustal method of DNASTAR software. Results IgM and IgG positive rate were 4.40% and 20.33% respectively for the crew from Southeast Asia country. The serum from clinical Dengue fever patients in Zhuhai in 2001 were inoculated into C6/36 Aedes albopictus cell line, and 1 strain of Dengue virus （ DV2 - 1 ） was isolated. The following identification confirmed to be dengue fever virus type 2 （ DEN - 2） by using a pair of universal primers and a dengue virus type 2 specific primer extend the certain sequence. The results showed that the isolated strain （DV2 - 1） was closely related to strain GD08/98, FJ11/99 and 16681. DV2 - 1 was more related to strain GD08/98, which was epidemic in Guangdong in 1998. Conclusion Dengue fever epidemic in Zhuhai in 2001 was caused Dengue virus type 2, which extrapolated the exotic infection.

关 键 词：登革病毒 病毒分离 序列分析 

分 类 号：R512.8[医药卫生—内科学]