中国兽医科学2024，Vol.54Issue(4)：492-497,6.DOI:10.16656/j.issn.1673-4696.2024.0061
鸡传染性喉气管炎TaqMan探针实时荧光定量PCR检测方法的建立与应用
Establishment and application of TaqMan probe real-time fluorescence quantitative PCR method for detection of infectious laryngotracheitis in chickens
摘要
Abstract
To establish a rapid diagnostic method for chicken infectious laryngotracheitis virus(ILTV),specific probes and primers were designed based on the ILTV gB gene,and a real-time fluorescent quantitative PCR(qPCR)method based on the TaqMan probe was established.The results showed that the standard curve of the ILTV qPCR method was y=-3.892 9x+44.607 with a linear correlation value(R2)of 0.998 8.The lowest detection limit of the method was 1.0× 10 copies/μL,and the coefficients of variation for both intra-and inter-batch reproducibility were less than 5.64％,and there was no cross-reactivity with other chicken-derived pathogens.The test results of the clinical samples showed a positive detection rate of 98.7％for the qPCR method compared to 92.61％for conventional PCR.The above results indicate that the method is sensitive,specific,and reproducible,and can be used for epidemic surveillance and epidemiologi-cal investigation of ILTV,providing a powerful tool for the prevention and diagnosis of ILTV infection.关键词
鸡传染性喉气管炎病毒/gB基因/实时荧光定量PCR/TaqMan探针Key words
chicken infectious laryngotracheitis virus/gB gene/real-time fluorescence quanti-tative PCR/TaqMan probe分类
农业科技引用本文复制引用
张相霞,王晓梦,张帅..鸡传染性喉气管炎TaqMan探针实时荧光定量PCR检测方法的建立与应用[J].中国兽医科学,2024,54(4):492-497,6.基金项目
江苏省科技项目(BE2021332,BE2022318) （BE2021332,BE2022318）
江苏省农业科技自主创新资金项目(CX(23)1004-3) （CX(23）
