Key Laboratory of Zoonosis of Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University, Beijing 100193, People's Republic of China.
World J Microbiol Biotechnol. 2013 Apr;29(4):607-16. doi: 10.1007/s11274-012-1216-x. Epub 2012 Nov 27.
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.
环介导等温扩增（LAMP）是一种新型基因扩增方法，本研究采用该方法检测猪呼吸道或肺部的支原体。在含有 10 fg 纯化靶 DNA（10 个支原体基因组拷贝）的 LAMP 反应中，病原体可在 30 分钟内被检测到，与实时 PCR 相当。在 63°C 下反应 30 分钟后，向 LAMP 反应体系中加入一定量的染料（SYBR Green I 和羟基萘酚蓝以适当的比例），通过目视检查可以轻松判断结果为阳性或阴性。此外，LAMP 能够区分支原体和其他密切相关的支原体菌株，表明具有高度的特异性。LAMP 检测更简单、更便宜，因为反应可以在等温条件下完成，并且需要的实验室基础设施更少。此外，该方法已被证明可用于现场鼻拭子和肺部样本中支原体的可靠诊断。
