Loop-mediated isothermal amplification for the rapid detection of Haemophilus parasuis

文献类型： 外文期刊

第一作者： Chen, Hao-Tai

作者： Chen, Hao-Tai;Chu, Yue-Feng;Liu, Yong-Sheng;Zhang, Jie;Lu, Zhong-Xin

作者机构：

关键词： Haemophilus parasuis;detection;loop-mediated isothermal amplification

期刊名称：FEMS IMMUNOLOGY AND MEDICAL MICROBIOLOGY （ 影响因子：3.078； 五年影响因子：2.747 ）

ISSN： 0928-8244

年卷期： 2010 年 60 卷 3 期

页码：

收录情况： SCI

摘要： Haemophilus parasuis infection is of considerable economic importance in the swine industry due to the high costs associated with treatment and loss of animals all over the world. In the present study, loop-mediated isothermal amplification (LAMP) is described for the rapid and specific detection of this species. A primer set derived from the inf B gene of H. parasuis was used to validate the assay using 15 H. parasuis reference strains, 39 clinical isolates, 75 positive samples, and 18 other pathogens. The results indicated that positive reactions were confirmed for all H. parasuis strains and specimens by LAMP after 45 min reaction at 65 degrees C in a water bath, and no cross-reactivity was observed from other non-H. parasuis strains. The detection limit of the conventional PCR was 25 copies, while that of the LAMP was five copies per tube. Therefore, the sensitivity of LAMP was higher than that of PCR. LAMP is likely to be more suitable as a routine diagnostic tool, especially in clinics without complicated equipment such as thermal cycling machines and electrophoresis apparatus. In these scenarios, the H. parasuis LAMP assay has the potential for field diagnosis.

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