现代畜牧科技Issue(3)：5-8,4.DOI:10.19369/j.cnki.2095-9737.2025.03.002
牛呼吸道病原体四重PCR检测方法的建立与应用
Establishment and Application of a Quadruple PCR Method for Detection of Bovine Respiratory Pathogens
摘要
Abstract
In order to establish one method for simultaneous detection of Bovine adenovirus virus type 3(BAdV-3),Bovine respiratory syncytial virus(BRSV),Mannheimia haemolytica(MH),and Bovine coronavirus(BCoV),four pairs of specific primers were designed according to the conserved sequences of BAdV-3、BRSV、MH and BCoV genes published by Genbank.The specificity,sensitivity and repeatability of the PCR were verified,and a quadruple PCR method was established to detect BAdV-3、BRSV、MH and BCoV at the same time,preliminary clinical application.The results showed that the optimal annealing temperature was 54℃and the optimal primer concentration was 0.5 μmol/L.The specific results showed that the quadruple PCR could only detect BAdV-3、BRSV、MH and BCoV.The minimum detection limits of BAdV-3、BRSV、MH and BCoV were 1.67×104、1.58×104、1.75×103、1.29×103copies/μL,respectively.The established quadruple PCR was used to detect 40 clinical samples,and the results were consistent with that of single PCR.The results show that the quadruple PCR method established in this study can be used for the epidemiological investigation of BAdV-3、BRSV、MH and BCoV.关键词
腺病毒/呼吸道合胞病毒/支原体/冠状病毒/四重PCRKey words
adenovirus/respiratory syncytial virus/mycoplasma/coronavirus/quadruple PCR分类
畜牧业引用本文复制引用
沈思思,王欢,王岩,刘文,冯万宇,陈亮,兰世捷,李丹,张蕾,周景明,刘雪松,张国华..牛呼吸道病原体四重PCR检测方法的建立与应用[J].现代畜牧科技,2025,(3):5-8,4.基金项目
黑龙江省农业科学院畜牧兽医分院2022年自拟项目(ZNKT-202213) （ZNKT-202213）
