Zhao Fei, Liu Zhong, Gu Yixin, Yang Yuelian, Xiao Di, Tao Xiaoxia, Meng Fanliang, He Lihua, Zhang Jianzhong
Chinese Center for Disease Control and Prevention, Beijing, China.
Acta Microbiol Immunol Hung. 2013 Mar;60(1):1-9. doi: 10.1556/AMicr.60.2013.1.1.
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.
肺炎支原体是引起儿童和成人呼吸道感染的最重要病原体之一。在本研究中，我们描述了一种快速灵敏的比色环介导等温扩增（LAMP）方法来检测肺炎支原体。用21种常见呼吸道病原体和39份肺炎支原体DNA检测了该检测方法的特异性和灵敏度。在39株肺炎支原体分离株中，LAMP的灵敏度为100%，在其他支原体的9个成员和12种常见呼吸道病原体中，特异性为100%。该检测方法的最低检测限（LDL）为102个拷贝，通过一系列标准肺炎支原体DNA进行检测。为评估LAMP检测方法的临床适用性，分别采用传统PCR、实时PCR和LAMP检测方法对80份临床样本进行了检测。阳性率分别为15.0%、32.5%和26.3%。这种比色LAMP检测方法在检测肺炎支原体方面显示出与传统PCR相当的高灵敏度。它是在中国农村地区和基层临床简单、经济有效且快速检测肺炎支原体的一种有价值的方法。
