中国动物检疫2024，Vol.41Issue(2)：72-79,8.DOI:10.3969/j.issn.1005-944X.2024.02.013
猪肺炎支原体SYBR Green I实时荧光定量PCR检测方法的建立与应用
Establishment and Application of SYBR Green I Real-time Fluorescence Quantitative PCR Assay for Mycoplasma hyopneumoniae
摘要
Abstract
In order to establish a rapid and efficient method to detect Mycoplasma hyopneumoniae(Mhp),4 pairs of specific primers were designed based on the conserved sequence of Mhp published by NCBI,and the optimal primer(SX4)was screened after correlation analysis based on the test results of the primers by fluorescence quantitative PCR and fitting curves to establish a Mhp SYBR Green I real-time fluorescence quantitative PCR assay.The method could be used to specifically detect Mhp,without specific amplification to porcine circovirus type 2,porcine parvovirus,pseudorabies virus,Haemophilus parasuis serotype 4 and Streptococcus suis serotype 2.The sensitivity of SX4 was up to 4.9×101 copies/μL,and inter/intra-group variable coefficient was less than 2%.The method could be used to detect the amount of Mhp from produced antigen and commercial vaccines.In conclusion,the Mhp SYBR Green I real-time fluorescence quantitative PCR assay was successfully established,and could be used to detect the amount of Mhp from produced antigen and clinical vaccines with the advantages of strong specificity,high sensitivity and good repeatability.关键词
猪肺炎支原体/荧光定量PCR/颜色改变单位(CCU)/相关性Key words
Mycoplasma hyopneumoniae/real-time fuorescent quantitative PCR/colour change unit(CCU)/correlation分类
农业科技引用本文复制引用
闫微,汤细彪,杨柳,龙云志,宋文博,李倩倩,余道兵,周明光,徐高原,黄超..猪肺炎支原体SYBR Green I实时荧光定量PCR检测方法的建立与应用[J].中国动物检疫,2024,41(2):72-79,8.基金项目
国家重点研发计划项目(2022YFD1800902) （2022YFD1800902）
