title: Development of a loop-mediated isothermal amplification targeting a gene within the pyruvate dehydrogenase complex, the pdhA gene, for rapid detection of Mycoplasma gallisepticum.
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.
authors.affiliation: Department of Poultry Infectious Diseases, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, China (Zhang, Bao, Yu, Cheng, Tan, Qiu, Song, Ding)College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu, China (Zhang, Fei)Poultry Institute, Chinese Academy of Agricultural Sciences, Yangzhou, Jiangsu, China (Dai).
authors.forename: Fanqing
authors.initials: F
authors.lastname: Zhang
authors.affiliation: Department of Poultry Infectious Diseases, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, China (Zhang, Bao, Yu, Cheng, Tan, Qiu, Song, Ding)College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu, China (Zhang, Fei)Poultry Institute, Chinese Academy of Agricultural Sciences, Yangzhou, Jiangsu, China (Dai).
authors.forename: Shijun
authors.initials: S
authors.lastname: Bao
authors.affiliation: Department of Poultry Infectious Diseases, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, China (Zhang, Bao, Yu, Cheng, Tan, Qiu, Song, Ding)College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu, China (Zhang, Fei)Poultry Institute, Chinese Academy of Agricultural Sciences, Yangzhou, Jiangsu, China (Dai).
authors.forename: Shengqing
authors.initials: S
authors.lastname: Yu
authors.affiliation: Department of Poultry Infectious Diseases, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, China (Zhang, Bao, Yu, Cheng, Tan, Qiu, Song, Ding)College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu, China (Zhang, Fei)Poultry Institute, Chinese Academy of Agricultural Sciences, Yangzhou, Jiangsu, China (Dai).
authors.forename: Jinghua
authors.initials: J
authors.lastname: Cheng
authors.affiliation: Department of Poultry Infectious Diseases, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, China (Zhang, Bao, Yu, Cheng, Tan, Qiu, Song, Ding)College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu, China (Zhang, Fei)Poultry Institute, Chinese Academy of Agricultural Sciences, Yangzhou, Jiangsu, China (Dai).
authors.forename: Lei
authors.initials: L
authors.lastname: Tan
authors.affiliation: Department of Poultry Infectious Diseases, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, China (Zhang, Bao, Yu, Cheng, Tan, Qiu, Song, Ding)College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu, China (Zhang, Fei)Poultry Institute, Chinese Academy of Agricultural Sciences, Yangzhou, Jiangsu, China (Dai).
authors.forename: Xvsheng
authors.initials: X
authors.lastname: Qiu
authors.affiliation: Department of Poultry Infectious Diseases, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, China (Zhang, Bao, Yu, Cheng, Tan, Qiu, Song, Ding)College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu, China (Zhang, Fei)Poultry Institute, Chinese Academy of Agricultural Sciences, Yangzhou, Jiangsu, China (Dai).
authors.forename: Cuiping
authors.initials: C
authors.lastname: Song
authors.affiliation: Department of Poultry Infectious Diseases, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, China (Zhang, Bao, Yu, Cheng, Tan, Qiu, Song, Ding)College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu, China (Zhang, Fei)Poultry Institute, Chinese Academy of Agricultural Sciences, Yangzhou, Jiangsu, China (Dai).
authors.forename: Yabin
authors.initials: Y
authors.lastname: Dai
authors.affiliation: Department of Poultry Infectious Diseases, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, China (Zhang, Bao, Yu, Cheng, Tan, Qiu, Song, Ding)College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu, China (Zhang, Fei)Poultry Institute, Chinese Academy of Agricultural Sciences, Yangzhou, Jiangsu, China (Dai).
authors.forename: Rongmei
authors.initials: R
authors.lastname: Fei
authors.affiliation: Department of Poultry Infectious Diseases, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, China (Zhang, Bao, Yu, Cheng, Tan, Qiu, Song, Ding)College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu, China (Zhang, Fei)Poultry Institute, Chinese Academy of Agricultural Sciences, Yangzhou, Jiangsu, China (Dai) shoveldeen@shvri.ac.cn.
authors.forename: Chan
authors.initials: C
authors.lastname: Ding
mesh_terms: D000818:Animals; D017931:DNA Primers; D009175:Mycoplasma Infections / Q000175:diagnosis / Q000662:veterinary*; D045686:Mycoplasma gallisepticum / Q000235:genetics / Q000302:isolation & purification*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods / Q000662:veterinary*; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary; D011200:Poultry; D011201:Poultry Diseases / Q000175:diagnosis* / Q000382:microbiology; D011237:Predictive Value of Tests; D011768:Pyruvate Dehydrogenase Complex / Q000235:genetics; D012680:Sensitivity and Specificity
chemical_list: D017931:DNA Primers; D011768:Pyruvate Dehydrogenase Complex
keywords: Detection; Mycoplasma gallisepticum; loop-mediated isothermal amplification