媒介动物(蚊、猪)体内黄病毒分子流行病学调查研究  

Molecule Epidemiologic Study for Vector-Born (Mosquito and Pig) Flavivirus in Zhuhai

作　　者：马洪波[1] 前田秋彦[2] 孙虹[1] 柯昌文[3] 杜坚[1] 谭华[1] 姚若东[1] 高岛郁夫[2] 柯明剑[1] 郑夔[3] 王遇春[1] 前田润子[2] 曾新宇[1] 冯子力[1] 

机构地区：[1]珠海出入境检验检疫局,珠海519010 [2]日本北海道大学研究院兽医学研究部 [3]广东省疾病预防控制中心,广州510300

出　　处：《中国国境卫生检疫杂志》2007年第5期267-272,共6页Chinese Journal of Frontier Health and Quarantine

基　　金：国家质量监督检验检疫总局科研基金项目:(2006IK179)

摘　　要：〔目的〕通过对珠海注册养猪场及珠海口岸地区的蚊虫体内黄病毒的带毒率、宿主动物(猪群)及其人群进行血清黄病毒类(日本脑炎、登革热等)抗体水平的调查研究,为控制黄病毒在人群中的流行提供科学依据。〔方法〕采集养猪场猪血清、养猪场和口岸蚊标本、养猪场从业人员、入境的东南亚船员和城区部分发热病人血清,采用酶联免疫吸附试验(ELISA)法分别检测猪群和人群血清中的日本脑炎、登革热抗体;使用紫外灯诱蚊法在养猪场和口岸捕捉蚊类,经鉴定种类后,按20只蚊一组制成蚊悬液后,采用细胞培养、逆转录-聚合酶链反应(RT-PCR)以及荧光定量PCR技术检测蚊体内日本脑炎病毒、登革和西尼罗病毒等黄病毒的携带情况。〔结果〕1.猪血清日本脑炎病毒IgG抗体:结果已发表于《中国国境卫生检疫杂志》2007年第3期《珠海市部分注册猪场日本脑炎调查研究》。2.蚊媒带毒情况检测:2005年1月～2006年12月共捕获的2763只成蚊,经鉴定隶属4属8种,其中白纹伊蚊所占比例最高(32.75%),其次为致倦库蚊(26.06%)、三带喙库蚊(25.30%)、海滨库蚊(7.75%)、中华按蚊(3.84%)、骚扰阿蚊(3.18%),其他(常型曼蚊和巨型阿蚊等)(1.12%);在132组蚊研磨液的病毒细胞培养中,有7份标本出现CPE病变,使用黄病毒通用引物,RT-PCR反应能扩增出相应的阳性条带,其中白纹伊蚊组阳性3份,致倦库蚊组阳性3份,三带喙库蚊组阳性1份,但用日本脑炎病毒、登革病毒Ⅰ～Ⅳ型和西尼罗病毒等特异性引物,未能扩增出相应的阳性条带,说明可能存在着其它黄病毒类的病毒感染;采用荧光定量PCR法,对蚊悬液和细胞培养液日本脑炎、登革和西尼罗病毒进行检测,从1组海滨库蚊标本中检出日本脑炎病毒核酸阳性,但强度较弱,这与珠海地区是乙脑低发地区相一致。3.人群登革热和日本脑炎血清抗体检测:从东南亚的入境船Objective To investigate Flavivirus in mosquito, serum antibody of Flavivirus （Japanese encephalitis, Dengue fever and so on） in host animal （pig） and human in Zhuhai hoggeries and Zhuhai port, hereby to provide scientifical evidences for strengthening monitoring, preventing and controlling the epidemic of Flavivirus. Methods Pig sera from hoggeries, human sera from Southeast Asian crew, fever patients in the urban city and workers in hoggeries were collected to detect serum antibodies for Japanese encephalitis virus （JEV） and Dengue virus （DV） by ELISA, mosquitoes were captured at 4 hoggeries and ports in Zhuhai from January 2005 to December 2006. Mosquitoes samples were sorted, identified to species, and were ground to make suspension which performed on groups containing 20 female mosquitoes per sample. RNA of virus were extracted directly from mosquito suspension, or after suspension was inoculated into cells of Aedes albopictus C6/36s and cultured. RT-PCR and Real-Time RT-PCR methods was carried out to detect JE,DV and West Nile virus （WNV） in mosquitoes. Results 1. Positive rate of Anti-JEV IgG antibodies in pigs： Data had been published in Chinese Frontier Health Quarantine Jun.2007,Vol 30,No.3 （Seroepidemiological Survey on Japanese Encephalitis at Pig Farms Registered in Zhuhai）. 2. Flavivirus in mosquitoes： 2 763 mosquitoes were captured together during January 2005 to December 2006. They belonged to 4 genera and 8 species, of which Aedes albopictus were the most dominant species, accouting for 32.75%, next was Culex fatigans （26.06%）, Culex tritaeniorhynchus （25.30%）, Culex Sitiens （7.75%）, Anopheles Sinensis （3.84%）, Armigeres subalbatus （3.18%）, other mosquito （1.12%）. 132 pools of mosquito suspensions were conducted by cell culture, 7 samples presented CPE pathological change. Culture supernatants were collected for the RT-PCR using Flavivirus universal primers, and 7 samples with CPE were amplified the amplicons produced the expected bands of 835 b

关 键 词：蚊 猪 黄病毒 带毒率 血清抗体 

分 类 号：R373.3[医药卫生—病原生物学]