中国畜牧兽医2016，Vol.43Issue(12)：3107-3113,7.DOI:10.16431/j.cnki.1671-7236.2016.12.004
猪链球菌2型实时荧光定量 PCR 检测方法的建立与应用
Establishment and Application of Quantitative Real-time PCR Method to Detect Streptococcus suis Serotype 2
摘要
Abstract
In this study,a quantitative Real-time PCR method using the specific primers according to CPS 2J gene was established to detect Streptococcus suis serotype 2.The result showed that the equation of standard curve was y=-3.073x+36.87,r =0.995,which demonstrated that the assay had good linear relationship.The melting curve analysis showed that there was only specific peak.Sensitivity test showed that the method could detect the template at the lowest concentra-tion of 1.0×10 1 copies/μL,which was 10 times higher than the ordinary PCR.The specific tests showed that this method could able to detect Streptococcus suis serotype 2 specially and had no <br> cross-reaction with other serotypes or other bacteria from swine.The CV of repeatability test was 0.37% to 0.63%,lower than 2.5%.The clinical diagnosis showed this assay was more sensitive than ordinary PCR and bacteria isolation.All the results showed that the established method was sensitive,specific and reproducible,which could be used for the rapid diagnosis and quantitative detection of Streptococcus suis serotype 2.关键词
猪链球菌 2 型/实时荧光定量 PCR/敏感性/特异性/重复性Key words
Streptococcus suis serotype 2/quantitative Real-time PCR/sensibility/specificity/re-peatability分类
农业科技引用本文复制引用
李建达,刘俊珍,杜以军,李俊,杨灵芝,王金宝,吴家强,于江,张玉玉,任素芳,陈蕾,郭立辉,孙文博,陈智,王淞..猪链球菌2型实时荧光定量 PCR 检测方法的建立与应用[J].中国畜牧兽医,2016,43(12):3107-3113,7.基金项目
泰山学者特聘专家工程经费 （）
山东省现代农业产业技术体系(SDAIT) （SDAIT）
山东省农业科学院重大科技成果培育计划(2014CGPY04) （2014CGPY04）
山东省农业重大应用技术创新项目 （）
山东省自然科学基金(ZR2015YL078) （ZR2015YL078）
山东省农业科学院青年科研基金(2015YQN51) （2015YQN51）
山东省农业科学院农业科技创新工程项目(CXGC2016B14) （CXGC2016B14）
