羊边界病毒纳米PCR检测方法的建立OA

[Objective]The aim is to develop a rapid detection method for border disease virus(BDV).[Method]Specific primers were designed based on the conserved 5'UTR sequence of BDV.By optimizing the reaction system parameters(including annealing temperature,primer concentration,and nanoparticle addition ratio),a BDV Nano-PCR detection method was successfully constructed.The sensitivity,specificity,and clinical applicability of this method were compared and analyzed and a parallel verification was conducted with those of the conventional PCR method.[Result]The optimized Nano-PCR could stably amplify a BDV-specific target of 210 bp,with a minimum detection limit of 1.1×10 copies/μL,and its sensitivity was 100 times higher than that of the conventional PCR(1.1×103 copies/μL).In the specific detection,this method showed no cross-reaction with bovine viral diarrhea virus,peste des petits ruminants virus,and Caprine enterovirus.The detection of 50 clinical samples showed that the positive detection rate of Nano-PCR was 6%(3/50),which was completely consistent with the results of the conventional PCR,but the sensitivity was significantly higher than that of the conventional PCR.[Conclusion]the established BDV Nano-PCR detection method has higher sensitivity and specificity than the conventional PCR,and can provide technical support for the early diagnosis and epidemiological monitoring of border disease virus infections.