畜牧兽医学报2024，Vol.55Issue(5)：2267-2272,6.DOI:10.11843/j.issn.0366-6964.2024.05.044
PEDV、PoRVA和PDCoV TaqMan三重RT-qPCR检测方法的建立与初步应用
Establishment and Preliminary Application of PEDV,PoRVA and PDCoV TaqMan Triple RT-qPCR Assay
摘要
Abstract
This paper aims to establish a triple fluorescence quantitative PCR method for the simultaneous detection of porcine epidemic diarrhea virus(PEDV),porcine rotavirus type A(PoRVA)and porcine deltacoronavirus(PDCoV).Primers and probes were designed for PEDV-N gene,PoRVA-VP6 gene and PDCoV-M gene,and their performance was evaluated after the conditions were optimized,and compared with commercial kits.The results showed that the triple RT-qPCR method established in this study had good specificity and did not amplify positive nucleic acids such as PRRSV,PCV2 and PRV.The detection limits of PEDV,PoRVA and PDCoV were all 1 copies·μL-1.The coefficient of variation within and between groups was less than 1％.The sample has wide adaptability,and the coefficient of variation is less than 1％when different sample types are detected.Compared with commercial kits,the detection coincidence rates of PEDV and PoRVA were 92.5％and 97.5％,and the detection range of PDCoV was wider,and the detection effect of PDCoV variant strains was still good.The detection method established in this study has the advantages of good specificity,high sensitivity,strong stability and wide sample adaptability,which provides a good detection method for clinical diarrhea samples.关键词
猪流行性腹泻病毒/猪A群轮状病毒/猪丁型冠状病毒/三重RT-qPCRKey words
porcine epidemic diarrhea virus/porcine rotavirus type A/porcine deltacoronavirus/triple RT-qPCR分类
农业科技引用本文复制引用
胡泽奇,李润成,谭祖明,谢秀艳,王江平,秦乐娟,李荣,葛猛..PEDV、PoRVA和PDCoV TaqMan三重RT-qPCR检测方法的建立与初步应用[J].畜牧兽医学报,2024,55(5):2267-2272,6.基金项目
湖南省重点研发计划项目(2023NK2017) （2023NK2017）
云南省重大科技专项计划(202202AE090032) （202202AE090032）
湖南省教育厅重点项目(21A0125) （21A0125）
湖南省普通高等学校科技创新团队支持计划资助 （）
