Zhao Fei, Zhang Jianzhong, Wang Xuemei, Liu Liyong, Gong Jie, Zhai Zhixiang, He Lihua, Meng Fanliang, Xiao Di
National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, State Key Laboratory of Infectious Disease Prevention and Control, Beijing 102206, China.
Intelligene Biosystems (Qingdao) Co., Ltd, Qingdao, China.
iScience. 2021 Apr 16;24(5):102447. doi: 10.1016/j.isci.2021.102447. eCollection 2021 May 21.
In this study, a multisite SNP genotyping and macrolide (ML) susceptibility gene test method for () was developed based on matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). The detection limit of this method for nucleic acids was 10 -10 copies/reaction. Six SNP site-based genotyping and 3 ML susceptibility sites could be detected simultaneously based on multiplex PCR and mass probe. Using the method constructed in this study, 141 Chinese clinical isolates were divided into 8 SNP types. All the SNP test results for the ML susceptibility gene were in line with those of the 23S rRNA sequencing results. With this method, the multisite SNP genotyping and ML susceptibility determination of can be completed simultaneously in one test, which greatly reduces the workload and cost, improves the genotyping ability of and deserves clinical application.
在本研究中，基于基质辅助激光解吸/电离飞行时间质谱（MALDI-TOF MS）开发了一种用于（）的多位点单核苷酸多态性（SNP）基因分型和大环内酯（ML）敏感性基因检测方法。该方法对核酸的检测限为10-10拷贝/反应。基于多重聚合酶链反应（PCR）和质量探针，可同时检测6个基于SNP位点的基因分型和3个ML敏感性位点。使用本研究构建的方法，将141株中国临床分离株分为8种SNP类型。ML敏感性基因的所有SNP检测结果均与23S rRNA测序结果一致。使用该方法，可在一次检测中同时完成（）的多位点SNP基因分型和ML敏感性测定，大大减少了工作量和成本，提高了（）的基因分型能力，值得临床应用。
