Detection of Mycoplasma pneumoniae by colorimetric loop-mediated isothermal amplification

Zhao, Fei and Liu, Zhong and Gu, Yixin and Yang, Yuelian and Xiao, Di and Tao, Xiaoxia and Meng, Fanliang and He, Lihua and Zhang, Jianzhong (2013) Detection of Mycoplasma pneumoniae by colorimetric loop-mediated isothermal amplification. Acta Microbiologica et Immunologica Hungarica, 60 (1). pp. 1-9. ISSN 1217-8950

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Abstract

Mycoplasma pneumoniae (M. pneumoniae) is one of the most important pathogens that cause respiratory tract infection in children and adults. In this study, we describe a rapid and sensitive colorimetric loop mediated isothermal amplification (LAMP) method to detect M. pneumoniae. The specificity and sensitivity of this assay were detected with 21 common respiratory pathogens and 39 M. pneumoniae DNA. The sensitivity of LAMP was 100% among 39 M. pneumoniae isolates and the specificity was 100% among 9 members of other Mycoplasma and 12 common respiratory pathogens. The lowest detectable limit (LDL) of this assay was 10² copies, which detected by a series of standard M. pneumoniae DNA. To evaluate the clinical applicability of the LAMP assay, a total of 80 clinical samples were examined by conventional PCR, real-time PCR and the LAMP assays, respectively. The positive rates were 15.0%, 32.5% and 26.3%, respectively. This colorimetric LAMP assay demonstrated a high level of sensitivity comparable with that of conventional PCR for the detection of M. pneumoniae. It is a valuable method for simple, cost-effective and rapid detection of M. pneumoniae in the rural areas and basic clinical of China.

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