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

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 gen...

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Bibliographic Details
Published in:Journal of veterinary diagnostic investigation 2015-05, Vol.27 (3), p.260-267
Main Authors: Zhang, Fanqing, Bao, Shijun, Yu, Shengqing, Cheng, Jinghua, Tan, Lei, Qiu, Xvsheng, Song, Cuiping, Dai, Yabin, Fei, Rongmei, Ding, Chan
Format: Article
Language:English
Subjects:
Animals
DNA Primers
Mycoplasma gallisepticum
Mycoplasma gallisepticum - genetics
Mycoplasma gallisepticum - isolation & purification
Mycoplasma Infections - diagnosis
Mycoplasma Infections - veterinary
Nucleic Acid Amplification Techniques - methods
Nucleic Acid Amplification Techniques - veterinary
Polymerase Chain Reaction - methods
Polymerase Chain Reaction - veterinary
Poultry
Poultry Diseases - diagnosis
Poultry Diseases - microbiology
Predictive Value of Tests
Pyruvate Dehydrogenase Complex - genetics
Sensitivity and Specificity
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Summary: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.
ISSN:1040-6387
1943-4936
DOI:10.1177/1040638715584155