畜牧与兽医2023，Vol.55Issue(12)：85-90,6.
非洲猪瘟病毒MGF-505R基因缺失株双重荧光RPA检测方法的建立及应用
Establishment and application of a double real-time RPA method for detection of MGF-505R gene-deleted mutants of African swine fever virus
摘要
Abstract
This study was to establish a rapid field screening method for screening African swine fever(ASF)gene-deleted mutants in or-der to identify the complex infection situation of ASF in China.Based on the epidemic trends of ASFV in China,the specific primers and exo probes were designed and synthesized by analyzing the conserved regions of the ASFV B646L and MGF-505R genes,in order to establish a double real-time RPA method for detection of the above-mentioned mutants.The sensitivity and specificity of the method were analyzed.In addition,the method was tested in clinical application using the optimized RPA.The result indicated that the double real-time RPA method could monitor the detection process within 15 min.Its minimum detection of RPA was up to 10 copies/response for both the genes.The meth-od was highly specific,without cross-reactions with classical swine fever virus,porcine respiratory and reproductive syndrome virus,pseud-orabies virus,porcine parvovirus and porcine circovirus type 2.Then,266 clinical samples were tested using the dual real-time RPA and WOAH recommended real-time PCR,with a coincidence rate of up to 100％.The double real-time RPA method for detecting ASFV MGF-505R gene-deleted mutants could be adapted to the mobile real-time RPA detector,which possessed the advantages of rapidity,high sensi-tivity and strong specificity.The method is applicable for rapid clinical detection of ASFV,and it could serve as a new screening method for prevention and control of ASF.关键词
非洲猪瘟/MGF-505R基因/B646L基因/双重荧光RPA/应用Key words
African swine fever/MGF-505R gene/B646L gene/dual real-time RPA/application分类
农业科技引用本文复制引用
邓俊花,吕继洲,陈冬杰,袁向芬,魏方,赵文军,吴绍强..非洲猪瘟病毒MGF-505R基因缺失株双重荧光RPA检测方法的建立及应用[J].畜牧与兽医,2023,55(12):85-90,6.基金项目
国家重点研发计划项目(2022YFD1800605) （2022YFD1800605）
