Yuan Xin, Bai Changqing, Cui Qian, Zhang Han, Yuan Jing, Niu Kaiwen, Feng Yuzhong, Jin Xin, Li Puyuan, Liu Huiying
Department of Respiratory and Critical Care Medicine, 307th Hospital of Chinese People's Liberation Army Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing City, China.
Medicine (Baltimore). 2018 Jun;97(25):e10806. doi: 10.1097/MD.0000000000010806.
To improve the diagnosis and treatment of Mycoplasma pneumoniae (Mp) infection and reduce the misuse of antibiotics, we sought to establish a loop-mediated isothermal amplification (LAMP) assay for rapid detection of Mp.Six primers specific for the Mp P1 gene were designed, and the LAMP method was used to rapidly detect Mp. The sensitivity of the LAMP method was determined by serial dilution of the standard Mp strain FH (standard strains of Mycoplasma pneumoniae). Specificity was assessed with 17 common pathogenic microorganisms in the respiratory tract. Patient samples were collected from the Department of Respiratory and Critical Care Medicine at the 307th Hospital of Chinese People's Liberation Army from March 2016 to May 2017, examined prospectively, and compared with diagnosis by quantitative real-time polymerase chain reaction (qRT-PCR).The LAMP assay for Mp detection can be completed within 60 minutes. The minimum detection limit was 39 pg/μL, and no cross-reaction was observed with 17 common respiratory tract pathogens. Of the 125 clinical specimens tested, 43 cases were positive by LAMP assay, and 40 cases were positive by qRT-PCR (P = .162). All 43 samples determined as positive by LAMP test were confirmed to be Mp by Mp P1 protein sequencing.The LAMP assay is suitable for rapid detection of Mp. It has high sensitivity and specificity, and the detection results are not inferior to those of qRT-PCR.
为提高肺炎支原体（Mp）感染的诊断和治疗水平，减少抗生素的滥用，我们试图建立一种环介导等温扩增（LAMP）检测方法用于快速检测Mp。设计了6条针对Mp P1基因的特异性引物，并采用LAMP方法快速检测Mp。通过对标准Mp菌株FH（肺炎支原体标准菌株）进行系列稀释来确定LAMP方法的灵敏度。用17种呼吸道常见致病微生物评估特异性。于2016年3月至2017年5月从中国人民解放军第307医院呼吸与危重症医学科收集患者样本，进行前瞻性检测，并与定量实时聚合酶链反应（qRT-PCR）诊断结果进行比较。用于检测Mp的LAMP检测可在60分钟内完成。最低检测限为39 pg/μL，与17种常见呼吸道病原体未观察到交叉反应。在检测的125份临床标本中，LAMP检测43例阳性，qRT-PCR检测40例阳性（P = 0.162）。LAMP检测确定为阳性的所有43份样本经Mp P1蛋白测序证实为Mp。LAMP检测适用于Mp的快速检测。它具有高灵敏度和特异性，检测结果不低于qRT-PCR。
