中国兽医科学2024，Vol.54Issue(4)：485-491,7.DOI:10.16656/j.issn.1673-4696.2024.0057
牛传染性鼻气管炎病毒和牛副流感病毒3型的双重荧光PCR方法的建立及应用
Establishment and application of a dual real-time PCR for simultaneous detection of infectious bovine rhinotracheitis virus and bovine parainfluenza virus type 3
摘要
Abstract
In order to establish a method for simultaneous detection of infectious bovine rhino-tracheitis virus(IBRV)and bovine parainfluenza virus type 3(BPIV3),a dual real-time PCR assay was successfully established by screening primers and probes,optimizing reaction system and conditions.This method could only detect IBRV and BPIV3 specifically,but other unrelated pathogens couldn't be detected;the coefficients of variation were less than 3％for both intra-group and inter-group;and detection limits of IBRV and BPIV3 plasmids were 15.2 copies/μL and 18.3 copies/μ L,respectively.Furthermore,a total of 143 samples from beef cattles with bovine respiratory disease complex(BRDC)collected from Sichuan,Chongqing,Hebei,Anhui and Jilin Provinces and 59 samples of yaks with BRDC collected from Sichuan and Qinghai were detected by the dual real-time PCR assay,and the results showed that the positive rates of IBRV and BPIV3 in beef cattles were 42.66％and 39.86％,respectively,and co-infection rate was 22.38％;while the positive rates of IBRV and BPIV3 in yaks were 44.07％and 30.51％,respectively,and the co-infection rate was 18.64％.The results in this study indicated that the dual real-time PCR assay established has the characteristics of high specificity,stability and sensitivity,making it suitable for the rapid detection of IBRV and BPIV3 from clinical samples,and the detection results enrich the epidemiological data of IBRV and BPIV3 in China.关键词
牛传染性鼻气管炎病毒/牛副流感病毒3型/双重荧光定量PCR/混合感染Key words
infectious bovine rhinotracheitis virus/bovine parainfluenza virus type 3/dual re-al-time PCR/co-infection分类
畜牧业引用本文复制引用
龚宏毅,汤承,岳华..牛传染性鼻气管炎病毒和牛副流感病毒3型的双重荧光PCR方法的建立及应用[J].中国兽医科学,2024,54(4):485-491,7.基金项目
"十四五"国家重点研发计划课题资助项目(2021YFD1600203) （2021YFD1600203）
国家农业产业技术体系四川肉牛创新团队专项资助项目(SCCXTD-2020-13) （SCCXTD-2020-13）
四川省高等学校重点实验室——动物医学实验室资助项目(2021PTJS34) （2021PTJS34）
