Development of a loop-mediated isothermal amplification targeting a gene within the pyruvate dehydrogenase complex, the pdhA gene, for rapid detection of Mycoplasma gallisepticum

Author:

Zhang Fanqing123,Bao Shijun123,Yu Shengqing123,Cheng Jinghua123,Tan Lei123,Qiu Xvsheng123,Song Cuiping123,Dai Yabin123,Fei Rongmei123,Ding Chan123

Affiliation:

1. Department of Poultry Infectious Diseases, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, China (Zhang, Bao, Yu, Cheng, Tan, Qiu, Song, Ding)

2. College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu, China (Zhang, Fei)

3. Poultry Institute, Chinese Academy of Agricultural Sciences, Yangzhou, Jiangsu, China (Dai)

Abstract

Mycoplasma gallisepticum infections impose a significant economic burden on the poultry industry. In the current study, a loop-mediated isothermal amplification (LAMP) assay was developed and optimized to detect M. gallisepticum based on a gene within the pyruvate dehydrogenase complex, the pdhA gene, which codes for the major subunit (E1α) in the complex. The reaction conditions were optimized, and the specificity was confirmed by successful amplification of several M. gallisepticum strains, while no amplification was detected with 20 other major bacterial and viral pathogens of poultry. Additionally, the LAMP assay achieved 10-fold higher sensitivity than an existing polymerase chain reaction (PCR) method. The LAMP assay was applied to swab samples collected from poultry farms and compared with PCR. The positive detection rate was 20.2% (37/183) by LAMP and 13.1% (24/183) by PCR. The LAMP assay could provide a cost-effective, quick, and sensitive method for the detection of M. gallisepticum.

Publisher

SAGE Publications

Subject

General Veterinary

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