中国动物传染病学报2024，Vol.32Issue(6)：81-88,8.
类NADC34 PRRSV TaqMan实时荧光定量PCR检测方法的建立
Development of a TaqMan Real-time Fluorescence Quantitative PCR for Detection of NADC34-Like PRRSV
摘要
Abstract
The geographical prevalence of NADC34-like PRRSV in China has been expanding since its first report in 2018 in China,which has evolved into a dominant strain in some regions.On account of no specific PCR or real-time PCR diagnostic method for NADC34-like PRRSV,conventional PRRSV typing and differential diagnosis methods may lead to misdiagnosis of such strains as NADC30-like PRRSV.Therefore,specific primers and probes were designed in this study based on the sequence differences between NADC30-like PRRSV and NADC34-like PRRSV in the Nsp2 gene,to develop a TaqMan fluorescent quantitative PCR assay.The results showed that the method developed in this study had good specificity and the positive standard plasmids at different dilutions had a good linear range from 2.8×101 copies/μL to 2.8×106 copies/μL with the R2 value of the standard curve 0.997.The assay had a sensitivity of 2.8×101 copies/μL,which showed it had a good sensitivity.The results also indicated that CV values of both intra-and inter-assay tests were less than 2.5%,showing a good reproducibility.In conclusion,this study successfully developed a real-time fluorescent quantitative PCR method for rapid detection of NADC34-like PRRSV,which provided a powerful tool for the investigation of the epidemic status of this strain in China.关键词
类NADC34 PRRSV/TaqMan探针/实时荧光定量PCRKey words
NADC34-like PRRSV/TaqMan probe/real-time PCR分类
农业科技引用本文复制引用
袁丽莉,朱振邦,刘盼娆,陈南华,李燕华,范娟,陈昌海,李向东..类NADC34 PRRSV TaqMan实时荧光定量PCR检测方法的建立[J].中国动物传染病学报,2024,32(6):81-88,8.基金项目
江苏省自然科学基金(BK20210804) （BK20210804）
