云南省文山中越边境地区虫媒病毒调查

Investigation on arboviruses at Sino-Vietnam border areas in Wenshan of Yunnan province

目的 对云南省文山县、河口县等中越边境地区开展虫媒病毒调查，以期了解当地蚊虫携带虫媒病毒情况。方法2007年9月在当地5个采集点共采集蚊虫8326只，包括库蚊6091只，中华按蚊1334只，刺扰伊蚊848只，阿蚊53只，并保存于液氮。经消毒、研磨、离心等处理后进行组织培养细胞接种，分离病毒和对病毒分离物进行血清学和分子生物学鉴定，以软件进行病毒的核苷酸序列比对和系统发生分析。结果从库蚊分离到4株对C6／36细胞致病变的病毒分离物，其他蚊种没有分离到病毒分离物。鉴定结果表明，标本WS0704-2与版纳病毒抗体反应，PCR鉴定为版纳病毒，该病毒基因组第12片段的进化关系同中国其他版纳病毒有明显差异，位于一条独立的进化枝中，在进化上相对独立。标本WS0704—1、WS0708—1、WS0708-2为浓核病毒。结论云南省文山中越边境地区存在多种蚊虫媒介并携带版纳病毒等虫媒病毒。

Objective To investigate arboviruses in Wenshan and Hekou county which are the Sino- Vietnam frontier regions of Wenshan, Yunnan province, China. Methods In September 2007, 6091 culicines,1334 anophelines, 848 aedes vexans and 53 armigeres obturbans were collected from 5 field sites. Mosquitoes were collected and stored in liquid nitrogen after classification. The mosquito pools were homogenized,and centrifuged,then the supematant was inoculated onto C6/ 36 and BHK-21 cells, and the viral isolates were identified by serological and molecular biological methods. Sequence alignment and phylogenetic analysis on the viral isolates were carried out using Clustal X 1.85, GENEDOC and MEGA4 software. Results A total of 4 pairs of virus isolated with C6/36 cells cytopathic effect were observed, and other mosquito species have not eytopathie effect. Strain WS0704-2 was Banna virus which identified by antibody response and PCR. Strain WS0704-1, WS0708-1, WS0708-2 were culex pipens pallens densovirus （CppDNV） which identified by PCR. The phylogenetic analysis the 12th segment showed significant difference between the new banna virus and other strains isolated in China. Conclusion There are many mosquito vectors in frontier regions （ China and Vietnam） of Wenshan in Yunnan province of China, and mosquito-borne arbovirus, such as BAV were isolated here.