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

作　　者：张相霞 王晓梦 张帅[2] ZHANG Xiangxia;WANG Xiaomeng;ZHANG Shuai(Animal Husbandry Development Promotion Centre,Pingyi County,Shandong Province,Linyi 273300,China;College of Animal Science and Technology,Shandong Agricultural University,Tai'an 271018,China)

机构地区：[1]山东省平邑县畜牧发展促进中心,山东临沂273300 [2]山东农业大学动物科技学院,山东泰安271018

出　　处：《中国兽医科学》2024年第4期492-497,共6页Chinese Veterinary Science

基　　金：江苏省科技项目(BE2021332,BE2022318);江苏省农业科技自主创新资金项目(CX(23)1004-3)。

摘　　要：为了建立一种鸡传染性喉气管炎病毒(ILTV)的快速诊断方法,基于ILTV g B基因设计特异性探针和引物,建立一种基于TaqMan探针的实时荧光定量PCR(qPCR)方法。结果显示,ILTV qPCR方法的标准曲线为y=-3.8929x+44.607,线性相关值(R^(2))为0.9988。该方法的最低检测限为1.0×10 copies/μL,批内与批间重复性检测的变异系数均小于5.64%,且与其他鸡源病原体无交叉反应。临床样本检测结果显示,该qPCR方法的检测阳性率为98.7%,而普通PCR的检测阳性率为92.61%。以上结果表明,该方法灵敏度高、特异性强、重复性好,可用于ILTV的疫病监测和流行病学调查,为ILTV感染的预防和诊断提供了有力工具。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=-3.8929x+44.607 with a linear correlation value(R^(2))of 0.9988.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 epidemiological 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 

分 类 号：S852..659.1[农业科学—基础兽医学]