安徽农业科学2011，Vol.39Issue(6)：3607-3609,3629,4.
副猪嗜血杆菌实时荧光PCR快速检测方法的建立和应用
Development of Real-Time Fluorescent PCR for Rapid Detection of Haempohlius parasuis
摘要
Abstract
[ Objective] To develop a real-time fluorescent PCR assay for rapid detection of Haempohlius parasuis (HPS). [ Method] According to the conservative sequences of 16 S rRNA genes of HPS published in G enBank,a pair of specific primers was designed. The real-time fluorescent PCR was developed by optimizing primer concentration and annealing temperature. And its specificity and reproducibility were evaluated. Ten HPS-saspected samples were detected by the developed method. [ Result] The lowest detecfion limit of the developed real-time fluorescent PCR was 50 copies/μl. This method had good reproducibility ,and its coefficient of variations was lower than 2％. Only HPS rather than Streptococcus suis type 2,Staphylococcus aureus,E. coli DH( ) ,and swine Salmonella typhi could be detected by the developed PCR. The HPS-positive samples detected by this method were also positive when they were detected by isolation or conventional PCR. [ Conclusion] The developed real-time fluorescent PCR is rapid,sensitive,specific and highly reproducible; thus,it can be used for repaid detection of HPS.关键词
副猪嗜血杆菌/实时荧光PCR/16 S rRNAKey words
Haempohlius parasuis/ Real-time fluorescent PCR/ 16 S rRNA分类
农业科技引用本文复制引用
李军,谢永平,潘艳,谢宇舟,禤雄标,陈泽祥,杨威,马春霞,胡帅,彭昊,许力干..副猪嗜血杆菌实时荧光PCR快速检测方法的建立和应用[J].安徽农业科学,2011,39(6):3607-3609,3629,4.基金项目
广西科技攻关项目(桂科攻0993009-1). （桂科攻0993009-1）
