作　　者： ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; (詹爱军）;

机构地区： 深圳出入境检验检疫局

出　　处： 《动物医学进展》 2013年第12期6-10,共5页

摘　　要： 鸡传染性法氏囊病(IBD)是危害我国养禽业的主要疫病之一,呈世界性分布,对养鸡业造成重大危害。在国际种禽贸易中,对IBDV检测是口岸检疫的主要对象之一。为了建立标准的鸡传染性法氏囊病病毒分子检测方法,采用世界动物卫生组织(OIE)推荐的带有通用引物和酶切位点的特异性引物,通过对6株不同毒株的反复试验,优化了针对A片段VP2基因的常规反转录-聚合酶链反应(RT-PCR)检测方法 (IBDV-VP2(604)-RT-PCR)和针对B片段VP1基因的RT-PCR检测方法(IBDV-VP1(642)-RT-PCR);针对A片段VP2基因设计特异性引物和探针,建立了优化IBDV特异性实时荧光定量反转录-聚合酶链反应(rRT-PCR)检测方法(IBDV-VP2-rRT-PCR)。将传染性法氏囊病疫苗-法倍灵(IBDV-BLEN)疫苗株进行系列稀释,两种RT-PCR检测方法的灵敏度均为10-3,而IBDV-VP2-rRT-PCR检测方法灵敏度为10-4。所建立的三种分子生物学检测方法特异,与其他家禽病毒无任何交叉反应。通过对自2007年以来18批221份田间样品检测表明,所建立的方法准确可靠,适合对鸡传染性法氏囊病病毒进行快速灵敏检测和监测。 Infectious bursal disease virus（ IBDV） is one of three severe acute diseases in China.IBDV had emerged worldwide and caused great loss to poultry production.And IBD is a quarantine disease in the avian and avian product trade.The rapid diagnosis is crucial to any control program of IBD.In order to construct a protocol of molecular diagnostic detection technique（ MDD） for IBDV,two conventional RT-PCR assays directed at the VP2gene in fragment A with 604bp production and VP1gene of in fragment B with 642bp respectively were optimized with the universal primers and IBDV-specific 3′extremity primers respectively.A Taqman real-time reverse transcription-PCR test IBDV（ IBDV-VP2-rRT-PCR） directed at the VP2gene in fragment A was developed specifically with 5strains.The sensitivities of three kinds of MDD assays were evaluated.The sensitivities of IBDV-VP2（ 604）-RTPCR and IBDV-VP1（ 642）-RT-PCR were equal in 10-3 dilution of IBDV-BLEN vaccine,and the sensitivity of IBDV-VP2-rRT-PCR was ten times superior to two conventional RT-PCR assays in 10-4 dilution.The specificity of three MDD assays developed had no cross reaction with others.These methods potentially allowed for more rapid,sensitive,and specific detection with 221 field samples for five years with monitoring and surveillance of IBDV.

关 键 词： 传染性法氏囊病 常规 实时荧光定量 快速检测

领　　域： [农业科学] [农业科学] [农业科学]