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Development of real-time fluorescent PCR for rapid detection of Haempohlius parasuis.

作者：

Jun Li;Yan Xie;Xuan XiongBiao;Chen ZeXiang;Yang Wei;Chao Ma;Hu Shuai;Hao Peng;Xu LiGan;Yan Xie;Yawen Pa

单位：

Guangxi Veterinary Research Institute, Nanning 530001, Chin;Guangxi Veterinary Research Institute, Nanning 530001, China

关键词：

hps;real-time fluorescent pcr;developed;method;detected;rapid detectio

摘要：

[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 GenBank, 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-suspected samples were detected by the developed method. [Result] The lowest detection limit of the developed real-time fluorescent PCR was 50 copies/μl. This method had good reproducibility, and its coefficient of variation was lower than 2%. Only HPS rather than Streptococcus suis type 2, Staphylococcus aureus, E. coli DH5 alpha, and swine Salmonella typhi could be detected by the developed real-time fluorescent PCR. The HPS-positive samples detected by this method were also positive when they were detected by isolation of bacteria or conventional PCR. [Conclusion] The developed real-time fluorescent PCR is rapid, sensitive, specific and highly reproducible; thus, it can be used for rapid detection of HPS.

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Development of real-time fluorescent PCR for rapid detection of Haempohlius parasuis.

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