牛传染性鼻气管炎病毒微滴式数字PCR检测方法的建立OA

To establish a detection method for IBRV with high sensitivity,high specificity,and absolute quantification capability,this study designed specific primers and probes targeting the gB gene of IBRV,thereby developing an IBRV droplet digital PCR(ddPCR)assay.Critical reaction parameters including annealing temperature and primer/probe concentrations were systematically optimized,followed by validation of the method's specificity,sensitivity,and concordance.The results demonstrated that:the assay exhibited excellent specificity,specifically detecting IBRV without any cross-reactivity with FMDV,BVDV,BRSV,BPIV 3.It showed high sensitivity,with a minimum detection limit as low as 0.94 copies/μL.It achieved a 100%concordance rate with the standard qPCR detection method,indicating high consistency.These findings confirm that the established IBRV ddPCR assay,characterized by strong specificity,high sensitivity,and excellent concordance,can meet the requirements for precise quantification of IBRV,early diagnosis,vaccine antigen quantification,import-export quarantine,and epidemiological surveillance.It thus provides technical support for the accurate detection of Infectious Bovine Rhinotracheitis.