中国兽医科学2024，Vol.54Issue(4)：479-484,6.DOI:10.16656/j.issn.1673-4696.2024.0076
猪传染性胃肠炎病毒TaqMan实时荧光定量PCR检测方法的建立与应用
Establishment and application of TaqMan real-time quantitative PCR for detection of porcine transmissible gastroenteritis virus
摘要
Abstract
In this study,we designed highly specific primers and probes for the N gene fragment of porcine transmissible gastroenteritis virus(TGEV),constructed the recombinant plasmid,and estab-lished the TaqMan fluorescence quantitative PCR detection method for the rapid clinical diagnosis and laboratory testing.The results showed that the method had good specificity.Compared with other porcine viruses,there was no cross-reaction between viruses with similar clinical symptoms.The Ct val-ues displayed good linear relationship in the range of 1×n109-1 × 101 copies/μL,R2=0.99.The lowest detection limit of this method was 10 copies/μL.The standard curve was y=-3.058 7x+37.7,and the slope was-3.058 7.The method has good repeatability,high sensibility,and strong specificity.The method is suitable for the detection of TGEV antigen nucleic acid in clinical samples and the study of the growth characteristics of laboratory viruses.It provides an effective test and detection tool for the accurate quantification and diagnosis of TGEV.关键词
猪传染性胃肠炎病毒/N基因/荧光定量PCR/TaqManKey words
transmissible gastroenteritis virus/Ngene/fluorescent quantitative PCR/TaqMan分类
农业科技引用本文复制引用
张羽欣,王树茂,段宏勇,赵款,董世山,高飞,李丽薇..猪传染性胃肠炎病毒TaqMan实时荧光定量PCR检测方法的建立与应用[J].中国兽医科学,2024,54(4):479-484,6.基金项目
上海市科技兴农项目(2022-02-08-00-12-F01162) （2022-02-08-00-12-F01162）
国家重点研发计划项目(2022YFD1800305,2022YFD1800804,2021YFD1801401) （2022YFD1800305,2022YFD1800804,2021YFD1801401）
中国农业科学院科技创新工程项目(CAAS-CSLPDCP-202301) （CAAS-CSLPDCP-202301）
