【摘要】 目的　利用细胞、分子生物学技术对贵州省自然界白纹伊蚊体内DEN带毒情况进行调查。方法　采集贵州省 9个地 (州 )市共计 18个县 (区 )白纹伊蚊幼虫标本 ,饲养为成蚊 ;制备蚊悬液 ,碘化钠法提取RNA ,用登革病毒(DEN)NS1基因区通用引物经逆转录 -聚合酶链反应 (RT -PCR)检测DEN核酸 ,限制性内切酶酶切鉴定阳性PCR产物并分型 ;蚊悬液接种C6 / 36细胞进行病毒分离 ,制作细胞片 ,间接免疫荧光法检测DEN抗原。结果　从独山县麻尾镇白纹伊蚊标本中分离到 1株DEN - 2型病毒 ,从凯里、黄果树和镇远 3处地方株白纹伊蚊体内检测出DEN核酸 ,用抗DEN1～ 4型单克隆抗体经间接免疫荧光 (IFA)和RT -PCR产物酶切分型鉴定分别为登革病毒 2型、4型。结论　贵州省存在DEN感染的自然循环更多还原

【Abstract】 Objective To study molecular epidemiology of dengue virus from Aedes albopictus field in Guizhou province.Methods 18 different population of Aedes albopictus larvals were collected in Guizhou.The total RNA from mosquito were extracted.The viral NS1 gene fragment were amplified with reverse transcriptase polymerase chain reaction(RT-PCR).And the products were digested with Mav I for typing.Dengue virus in mosquito was isolated with C6/36 cells.Then the viral antigen was detected by indirect immunofluorescence assay(IFA).Results One strain of dengue virus was isolated with C6/36 cells from field Aedes albopictus in Mawei town.It was identified as dengue virus type 2 by IFA with monoclonal antibody and RT-PCR.Dengue virus type 2 or 4 RNA were detected in Aedes albopictus field collected from Kaili,Huangguoshu and Zhenyuan areas in Guizhou,respectively.Conclusion These findings highlighted the existence of a silent epidemic in Guizhou.更多还原