CAJ | 학술논문

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。
계전염성법씨낭병（IBD）시위해아국양금업적주요역병지일，정세계성분포，대양계업조성중대위해。재국제충금무역중，대 IBDV 검측시구안검역적주요대상지일。위료건립표준적계전염성법씨낭병병독분자검측방법，채용세계동물위생조직（OIE）추천적대유통용인물화매절위점적특이성인물，통과대6주불동독주적반복시험，우화료침대 A 편단 VP2기인적상규반전록-취합매련반응（RT-PCR）검측방법（IBDV-VP2（604）-RT-PCR）화침대 B 편단 VP1기인적 RT-PCR 검측방법（IBDV-VP1（642）-RT-PCR）；침대 A 편단 VP2기인설계특이성인물화탐침，건립료우화 IBDV 특이성실시형광정량반전록-취합매련반응（rRT-PCR）검측방법（IBDV-VP2-rRT-PCR）。장전염성법씨낭병역묘-법배령（IBDV-BLEN）역묘주진행계렬희석，량충 RT-PCR 검측방법적령민도균위10-3，이 IBDV-VP2-rRT-PCR 검측방법령민도위10-4。소건립적삼충분자생물학검측방법특이，여기타가금병독무임하교차반응。통과대자2007년이래18비221빈전간양품검측표명，소건립적방법준학가고，괄합대계전염성법씨낭병병독진행쾌속령민검측화감측。
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 mo-lecular diagnostic detection technique(MDD)for IBDV,two conventional RT-PCR assays directed at the VP2 gene in fragment A with 604bp production and VP1 gene of in fragment B with 642 bp respectively were optimized with the universal primers and IBDV-specific 3′extremity primers respectively.A Taqman real-time reverse transcrip-tion-PCR test IBDV(IBDV-VP2-rRT-PCR)directed at the VP2 gene in fragment A was developed specifically with 5 strains.The sensitivities of three kinds of MDD assays were evaluated.The sensitivities of IBDV-VP2(604)-RT-PCR and IBDV-VP1(642)-RT-PCR were equal in 10-3 dilution of IBDV-BLEN vaccine,and the sensitivity of IB-DV-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.