线性探针法用于分枝杆菌菌种快速鉴定的应用研究

doi:10.19871/j.cnki.xfcrbzz.2022.02.010

摘要/Abstract

摘要： 目的探讨线性探针法用于分枝杆菌菌种快速鉴定的可行性和应用价值。方法收集2017年1月至2018年12月深圳市慢性病防治中心、光明区人民医院和盐田区人民医院登记疑似结核病患者的分枝杆菌阳性培养物3420份。经GenoType MTBDRplus结核分枝杆菌耐药检测试剂盒初筛和剔除患者的重复样本后,筛选出127株非结核分枝杆菌（NTM）进行菌种鉴定。采用线性探针法和PCR基因测序法（16S rRNA和ITS）对NTM菌株进行菌种鉴定,并以测序结果为菌种鉴定的“金标准”。结果 127株NTM的菌种分布为：脓肿分枝杆菌47株（37.0%）,偶发分枝杆菌20株（15.8%）,胞内分枝杆菌18株（14.2%）,戈登分枝杆菌11株（8.7%）,堪萨斯分枝杆菌8株（6.3%）,鸟-胞内分枝杆菌复合群7株（5.5%）,慢生黄分枝杆菌6株（4.7%）,奇美拉分枝杆菌2株（1.6%）,龟分枝杆菌1株（0.8%）,蟾分枝杆菌1株（0.8%）,瘰疬分枝杆菌1株（0.8%）,外来分枝杆菌1株（0.8%）,土分枝杆菌1株（0.8%）,美容品分枝杆菌1株（0.8%）,熊本分枝杆菌1株（0.8%）和富养二氧分枝杆菌1株（0.8%）。与PCR相比,线性探针法对127株NTM全部检出,其中112株（112/127,88.2%）能准确鉴定到菌种,2株（2/127,1.6%）不能区分为瘰疬/胞内分枝杆菌,12株（12/127,9.4%）超出试剂盒鉴定范围,1株外来分枝杆（1/127,0.8%）鉴定错误。结论线性探针法可以快速、准确地鉴定常见的分枝杆菌菌种,但菌种鉴定范围有限,不能完全满足临床菌种鉴定的需求。

关键词: 非结核分枝杆菌, 线性探针法, 基因测序, 鉴定

Abstract: Objective To study and evaluate the GenoType Mycobacterium CM for rapid identification of NTM. Method A total of 3420 Mycobacterium positive cultures were collected from 2017 to 2018 in Shenzhen Center for Chronic Disease Control, Guangming District People's Hospital and Yantian District People's Hospital. After initial detection of GenoType MTBDRplus and elimination of the duplicate samples of patients, 127 cultures were screened out for further NTM identification through GenoType Mycobacterium CM and PCR sequencing (16S rRNA and ITS). The PCR sequencing results were used as the "gold standard" to evaluate the results of GenoType Mycobacterium CM. Result Among 127 NTM strains, there were 47 M.abscessus strains (37.0%), 20 M.fortuitum strains (15.8%), 18 M.intracellularstrains (14.2%), 11 M.gordonae strains (8.7%), 8 M.kansasii strains (6.3%), 7 M.avium strains (5.5%), 6 M.lentiflavum strains (4.7%), and 2 M.chimaera strains (1.6%), 1 M.xenopi strain (0.8%), 1 M.scrofulaceum strain (0.8%), 1 M.peregrinum strain (0.8%), 1 M.terraestrain (0.8%), 1 M.cosmeticum strain (0.8%), 1 M.kumamotonense strain (0.8%) and 1 M.dioxanotrophicus strain (0.8%). Compared with sequencing results, all 127 strains of NTM were detected by GenoType Mycobacterium CM, and 112 strains (112/127, 88.2%) could be accurately identified, 2 strains (2/127, 1.6%) could not be distinguished as M.intracellular or M.intracellular, 12 strains (12/127, 9.4%) were beyond the identification range of the kit, and 1 M.peregrinum strain (1/127, 0.8%) was incorrectly identified. Conclusion GenoType Mycobacterium CM can quickly and accurately identify common NTM, but its scope of strain identification is limited and can not fully meet the needs of clinical NTM identification.

Key words: Non-tuberculous mycobacterium, Line probe, Gene sequencing, Identification

朱玉梅, 钟育权, 李金莉, 吴利凯, 王峰. 线性探针法用于分枝杆菌菌种快速鉴定的应用研究[J]. 新发传染病电子杂志, 2022, 7(2): 47-51.

Zhu Yumei, Zhong Yuquan, Li Jinli, Wu Likai, Wang Feng. Evaluation of GenoType Mycobacterium CM for the rapid identification of non-tuberculous mycobacteria[J]. Electronic Journal of Emerging Infectious Diseases, 2022, 7(2): 47-51.

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