Detection of Haemophilus parasuis isolates from South China by loop-mediated isothermal amplification and isolate characterisation

文献类型： 外文期刊

第一作者： Zhang, Jian-min

作者： Zhang, Jian-min;Shen, Hai-yan;Liao, Ming;Ren, Tao;Guo, Li;Xu, Cheng-gang;Feng, Sai-xiang;Fan, Hui-ying;Li, Jing-yi;Chen, Ji-dang;Zhang, Bin;Shen, Hai-yan

作者机构：

期刊名称：ONDERSTEPOORT JOURNAL OF VETERINARY RESEARCH （ 影响因子：1.792； 五年影响因子：2.113 ）

ISSN： 0030-2465

年卷期： 2012 年 79 卷 1 期

页码：

收录情况： SCI

摘要： Haemophilus parasuis is the etiological agent of Glasser's disease, which is characterised by fibrinous polyserositis, meningitis and polyarthritis, causing severe economic losses to the swine industry. In this study, a loop-mediated isothermal amplification (LAMP) test was developed to improve the specificity, facility and speed of diagnosis of H. parasuis isolates. The LAMP assay rapidly amplified the target gene within 50 min incubation at 63 C in a laboratory water bath. The LAMP amplicon could be visualised directly in the reaction tubes following the addition of SYBR Green I dye. The detection limit of this LAMP method was 10 CFU/mL, which was 10 times more sensitive than the earlier 16S rRNA polymerase chain reaction (PCR) test conducted by Oliveira, Galina and Pijoan (2001), and no cross-reactivity was observed from other non-H. parasuis strains. This LAMP test was evaluated further on 187 clinical specimens from pigs suspected of being infected with H. parasuis. Forty-three were found positive by bacterial isolation of H. parasuis, as well as by the 16S rRNA PCR and LAMP tests. The 43 H. parasuis isolates were classified into 9 serovars and had 37 genetic patterns when analysed by pulsed-field gel electrophoresis (PFGE). This displayed that various H. parasuis serovars and genotypes were widely distributed in South China. Therefore, the speed, specificity and sensitivity of the LAMP test, the lack of a need for expensive equipment, and the visual readout showed great potential for a correct clinical diagnosis of H. parasuis in favour of controlling Glasser's disease.

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