鸡黄病毒RT-PCR检测方法的建立  

Establishment of RT-PCR for detection of chicken flavivirus

作　　者：王劭[1,2] 陈仕龙[1,2] 陈少莺[1,2] 程晓霞[1,2] 林锋强[1,2] 朱小丽[1] 江斌[1] 李兆龙[1] 

机构地区：[1]福建省农业科学院畜牧兽医研究所,福建福州350013 [2]福建省畜禽疫病防治工程技术研究中心,福建福州350013

出　　处：《中国预防兽医学报》2012年第7期548-550,共3页Chinese Journal of Preventive Veterinary Medicine

基　　金：福建省农科院创新团队项目(STIF-Y02);福建省公益类科研院所专项(2011R1025-2)

摘　　要：为建立鸡黄病毒(CFV)分子生物学检测方法,本研究以虫媒介黄病毒基因3'末端通用引物EMF1/VD8作为鉴别引物,以CFV分离株CJD-05为模板,建立了检测CFV的RT-PCR方法。RT-PCR扩增产物测序结果显示,其扩增片段为708 bp,而该方法对其它禽类病毒病原核酸的扩增结果均为阴性,表明该方法具有较高的特异性。扩增的特异性片段与GenBank中CFV、鸭黄病毒和鹅黄病毒序列同源性达到100%、98%和99%。对参考病毒进行梯度稀释检测,该方法检测CFV的敏感度可达10-3TCID50/0.1 mL。采用所建立的RT-PCR方法对2009年福建省部分鸡场的112份临床样品进行检测,结果显示37份为CFV阳性。本研究建立的RT-PCR方法可以作为CFV在临床检测和分子流行病学调查的一种检测方法。To establish a RT-PCR based detection for chicken flavivirus （CFV） infection, , a pair of arthropod-transmitted flavivi^s universal primers of EMF1 and VD8 in accordance with 3＇ terminal sequence was used to amplify target sequence from the RNA of CFV, and a special fragment of 708 bp was amplified from CFV CJD-05 with a detection limit of 10-3 TCIDs0, but no amplification was found from other related avian viruses. Sequence analysis showed that the amplified 3＇ terminal fragment shared 100%, 98%, 99% identity with the CFV, duck flavivirus and goose flavivims, respectively. The RT-PCR was applied to detect 112 clinical samples from several chicken farms of Fujian province in 2009, and 33% （37/112） samples were CFV-positive. The result showed that the RT-PCR technique provided a useful method for clinical diagnosis and epidemiologic survey of CFV.

关 键 词：鸡黄病毒 RT-PCR 建立 应用 

分 类 号：S852.65[农业科学—基础兽医学]