生物灾害科学2025，Vol.48Issue(3)：354-362,9.DOI:10.3969/j.issn.2095-3704.2025.03.45
猪圆环病毒2型、3型、4型三重PCR检测方法的建立与初步应用
The construction and preliminary application of triplex PCR to the simultaneous detection of Porcine circovirus type 2,Porcine circovirus type 3 and Porcine circovirus type 4
摘要
Abstract
[Objective]This study aimed to construct a low-cost and rapid detection method for porcine circovirus type 2(PCV2),porcine circovirus type 3(PCV3)and porcine circovirus type 4(PCV4).[Method]A triplex PCR was developed based on three pairs of specific primers for each of PCV2,PCV3 and PCV4 Rep gene,and a triple PCR method for the simultaneous detection of the above three circoviruses was established by optimizing the reaction conditions.[Result]Results showed that the established triple PCR method could efficiently amplify the genomic DNA of PCV2,PCV3 and PCV4,but not specifically amplify the nucleic acids of porcine reproductive and respiratory syndrome virus(PRRSV),classical swine fever virus(CSFV),porcine pseudorabies virus(PRV),porcine epidemic diarrhea virus(PEDV),porcine deltacoronavirus(PDCoV),porcine parvo virus(PPV),Transmissiblegast roenteritisof swine virus(TEGV)and Senecavirus A(SVA),with strong specificity;the minimum detection limits of recombinant plasmids for PCV2,PCV3 and PCV4 were 3.5×103 copies/μL,5.6×103 copies/μL,and 4.7×103 copies/μL,respectively,which were consistent with the single PCR detection results established by this experimenter and had high detection sensitivity.The results also showed that the positive rate of PCV2 was 64%(16/25),that of PCV3 32%(8/25),and that of PCV4 40%(10/25).The incidence of co-infection(PCV2/3,PCV2/4,PCV3/4)was 8%(2/25),while that of triple co-infection was 4%(1/25).The results of the single PCR primer in this test were consistent.[Conclusion]The triple PCR method established in this study can effectively detect PCV2/3/4 at the same time,which provides technical support for the detection of porcine circovirus co-infection and the epidemiological investigation of the disease.关键词
猪圆环病毒2型/猪圆环病毒3型/猪圆环病毒4型/三重PCRKey words
PCV2/PCV3/PCV4/triplex PCR分类
农业科技引用本文复制引用
梁惠平,喻富伟,尹经毅,张俊杰,吴欢生,吴虹瑾,肖艳,曾庆华,刘琪,黄曼孜,李昕宇,陈翰卓,文姝寓..猪圆环病毒2型、3型、4型三重PCR检测方法的建立与初步应用[J].生物灾害科学,2025,48(3):354-362,9.基金项目
江西省现代农业产业体系项目(JXARS-09) （JXARS-09）
