【摘要】 为调查我国部分地区规模化养鸡场传染性法氏囊病病毒(IBDV)新型变异株流行现状，参考IBDV VP2蛋白高变区序列设计Taqman-MGB探针和引物，建立IBDV快速检测方法，利用其检测来自河北邯郸、江西宜春、重庆7家大型育雏场送检法氏囊病料，并对阳性病料进行病毒分离、测序和进化分析。成功建立了一种兼顾IBDV鉴定和测序分型的荧光定量PCR方法，该方法在10~1～10~8拷贝/μL范围内具有良好线性关系(相关系数为0.9963),扩增效率良好(扩增效率为109%),对其他常见病毒无扩增，最低检测限为1.25×10~1拷贝/μL;利用该方法检测邯郸地区4家育雏场送检样品均为阳性，测序均为IBDV新型变异株，采用鸡胚自阳性样品中分离到3株IBDV(T2021与T2022、J2021);对分离株A节段测序发现，3株IBDV分离株均属国内新型变异株，与国内新型变异株SHG19氨基酸同源性为100%,核苷酸同源性为98.0%～98.6%,T2021与T2022、J2021分属两个进化分支。建立了一种IBDV快速检测方法，发现IBDV国内新型变异株在我国部分地区仍存在。更多还原

【Abstract】 The aim of this study was to investigate the prevalence of a new variant of infectious bursal disease virus in large-scale chicken farms in some regions of China.Taqman-MGB probe and primers were designed based on the hypervariable region sequence of IBDV VP2 protein, and a rapid detection method for IBDV was established.The samples were collected from seven large brood sites in Handan of Heibei province, Yichun of Jiangxi province and Chongqing, and the positive samples were isolated, sequenced and analyzed for virus evolution.Results showed that a fluorescence quantitative PCR method was successfully established, which was suitable for both IBDV identification and sequencing typing.The method had good linearity in the range of 10~1-10~8 copies/μL(correlation coefficient is 0.9963),and the amplification efficiency is 109%.The minimum detection limit is 1.25×10~1 copies/μL.By using this method, all the samples from four breeding farms in Handan were positive, all the sequences are the novel antigen variant strains IBDV.Three IBDV strains(T2021,T2022,J2021) were isolated from the positive samples of chicken embryos.Segment A sequencing showed that the three IBDV isolates are all novel antigen variant strains IBDV.The amino acid homology and nucleotide homology are 100% and 98.0%-98.6% with SHG19,respectively.T2021,T2022 and J2021 belong to two evolutionary branches.This study established a rapid detection method for IBDV,and found the novel antigen variant strains IBDV existing in some areas of China.更多还原