福建农业学报2025，Vol.40Issue(4)：370-376,7.DOI:10.19303/j.issn.1008-0384.2025.04.006
猪A群轮状病毒TaqMan荧光定量RT-PCR检测方法的建立与运用
A TaqMan RT-qPCR Assay for Detecting Porcine Rotavirus Group A
摘要
Abstract
[Objective]A rapid method for detecting porcine rotavirus group A(PoRV A)in disease monitoring and epidemiological investigation was developed.[Method]Specific primers and probes were selected based on the VP6 gene sequences of PoRV A in GenBank(accession numbers MT025937.1,OP978242.1 and PP566178.1).Reaction conditions of the Taq Man RT-qPCR method were optimized followed by evaluations on assay specificity,sensitivity,repeatability,and clinical application.[Results]The newly developed method could specifically amplify PoRV nucleic acid with a minimum detection limit of 27.0 copies·μL-1 and a sensitivity 100 times higher than RTPCR.There was no cross reactivity with the nucleic acids of PEDV,TGEV or PDCoV.The method showed high repeatability with the variation coefficients on intra-and inter-group below 1.10%.On 151 clinical specimens suspected of PoRV,a detection rate of 42.38%(64/151),which was better than that of conventional RT-PCR at 33.11%(50/151),was obtained by the assay.[Conclusion]The new Taq Man RT qPCR method for detecting VP6 gene of PoRV A was high in assay sensitivity,specificity,and repeatability.It was considered suitable for PoRV detection and epidemiological investigations.关键词
猪A群轮状病毒/TaqMan探针/荧光定量RT-PCRKey words
porcine rotavirus Group A/TaqMan probe/RT-qPCR分类
农业科技引用本文复制引用
林青,康龙滨,吴瑞森,赵文娟,陈秋勇,王隆柏,周伦江,俞道进..猪A群轮状病毒TaqMan荧光定量RT-PCR检测方法的建立与运用[J].福建农业学报,2025,40(4):370-376,7.基金项目
福建省农业高质量发展超越"5511"协同创新工程项目(XTCXGC2021008) （XTCXGC2021008）
福建省农业科学院科技创新团队建设项目(CXTD2021007-2) （CXTD2021007-2）
