title: Loop-mediated isothermal amplification for rapid and convenient detection of Mycoplasma hyopneumoniae.
abstract: Loop-mediated isothermal amplification (LAMP), a novel method of gene amplification, was employed in this study for detecting Mycoplasma hyopneumoniae in the respiratory tract or lungs of swine. The pathogen can be detected in LAMP reactions containing as few as 10 fg purified target DNA (10 copies of M. hyopneumoniae genome) within 30 min, which was comparable to real-time PCR. After 30-min reaction at 63 °C, the addition of a certain amount of dye (SYBR Green I and hydroxyl naphthol blue at a proper ratio) into the LAMP reaction system makes the results easily determined as positive or negative by visual inspection. In addition, the LAMP was able to distinguish between M. hyopneumoniae and other closely-related mycoplasma strains, indicating a high degree of specificity. The LAMP assay was more simple and cheap, since the reaction could be completed under isothermal conditions and less laboratorial infrastructure are required. And, it was proven reliable for M. hyopneumoniae diagnosis of nasal swab and lung samples from the field.
authors.affiliation: Key Laboratory of Zoonosis of Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University, Beijing 100193, People's Republic of China.
authors.forename: Jiahe
authors.initials: J
authors.lastname: Li
authors.forename: F Chris
authors.initials: FC
authors.lastname: Minion
authors.forename: Andrew C
authors.initials: AC
authors.lastname: Petersen
authors.forename: Fei
authors.initials: F
authors.lastname: Jiang
authors.forename: Sheng
authors.initials: S
authors.lastname: Yang
authors.forename: Panpan
authors.initials: P
authors.lastname: Guo
authors.forename: Jinxiang
authors.initials: J
authors.lastname: Li
authors.forename: Wenxue
authors.initials: W
authors.lastname: Wu
mesh_terms: D000818:Animals; D001431:Bacteriological Techniques / Q000191:economics / Q000379:methods*; D025202:Molecular Diagnostic Techniques / Q000191:economics / Q000379:methods*; D045705:Mycoplasma hyopneumoniae / Q000235:genetics / Q000302:isolation & purification*; D021141:Nucleic Acid Amplification Techniques / Q000191:economics / Q000379:methods*; D045729:Pneumonia of Swine, Mycoplasmal / Q000175:diagnosis*; D012680:Sensitivity and Specificity; D013194:Staining and Labeling / Q000379:methods; D013552:Swine; D013696:Temperature; D013997:Time Factors; D014730:Veterinary Medicine / Q000191:economics / Q000379:methods*