微阵列芯片法和多同源基因序列测序在非结核分枝杆菌菌种鉴定中的差异  

Difference in identification of non-tuberculous Mycobacterium between microarray chip and multiple homologous sequence

作　　者：郭明日[1] 李玉明[1] 李妍[1] 孙海柏[1] GUO Mingri;LI Yuming;LI Yan;SUN Haibai(Department of Clinical Laboratory,Tianjin Haihe Hospital,Tianjin University Haihe Hospital,China Key Research Laboratory for Infectious Disease Prevention for State Administration of Traditional Chinese Medicine,Tianjin 300350,China)

机构地区：[1]天津市海河医院、天津大学海河医院检验科、国家中医药管理局传染病重点研究室,天津300350

出　　处：《检验医学》2023年第10期909-914,共6页Laboratory Medicine

基　　金：天津市卫生健康科技项目(ZC20102);天津市津南区科技局科技项目(201805003)。

摘　　要：目的比较微阵列芯片法和多同源基因序列测序在非结核分枝杆菌(NTM)菌种鉴定中的差异,为NTM感染提供精准诊断依据。方法采用罗氏固体培养法复苏天津市海河医院2016—2019年保存的170株NTM菌株,分别采用微阵列芯片法和同源基因序列[16S rRNA、RNA聚合酶亚基B(rpoB)、热休克蛋白65(hsp65)]测序进行鉴定。以多同源序列测序分析结果为参考,分析微阵列芯片法菌种鉴定的准确性。比较2种方法样本周转时间(TAT)的差异。结果微阵列芯片法鉴定出8个NTM菌种,同源序列测序鉴定出14个。微阵列芯片法无法鉴定出缓慢生长群的猿分枝杆菌、慢生黄分枝杆菌、副瘰疬分枝杆菌和快速生长群的龟分枝杆菌、马德里分枝杆菌、母牛分枝杆菌等10个菌种,多同源序列测序可鉴定出全部的菌种。以多同源序列测序分析结果为参考,微阵列芯片法鉴定错误7株,鉴定到复合群的正确率为90.0%(153/170);16S rRNA、rpoB、hsp65单独和3个同源序列联合分析鉴定到复合群的正确率分别为93.5%、98.8%、99.4%和100.0%。对于微阵列芯片法无法区分的53株龟/脓肿分枝杆菌复合群(MABC),3个同源序列联合测序鉴定出6株龟分枝杆菌、25株MABC脓肿亚种、20株MABC马赛亚种和2株MABC bolletii亚种。微阵列芯片法单个样本鉴定TAT为8 h,170株NTM菌株全部鉴定需要2 d;多同源序列测序单个样本鉴定TAT为3 d,170株NTM菌株全部鉴定约需要5 d。结论微阵列芯片技术可快速鉴定临床常见分枝杆菌。多同源基因序列测序不仅可鉴定临床常见分枝杆菌,还可鉴定微阵列芯片法无法鉴定出的菌种和可能鉴定错误的菌种及其亚种,但所需时间较微阵列芯片法长。Objective To compare microarray chip and multiple homologous sequence in the identification of non-tuberculous Mycobacterium(NTM),and to provide accurate diagnosis reference for the clinical diagnosis and treatment of NTM infection.Methods For preserved NTM from 2016 to 2019 in Tianjin Haihe Hospital,using microarray chip and multiple homologous sequence,including 16S rRNA,subunit of RNA polymerase B(rpoB),heat shock protein 65(hsp65),the 170 isolates of NTM were analyzed.The accuracy of microarray chip was analyzed for NTM using the identification results of multiple homologous sequence as reference method.The turn-around times(TAT)were compared between the 2 methods.Results Totally,8 NTM species were identified by microarray chip,and 14 NTM species were identified by multiple homologous sequence.Microarray chip failedto identify 10 isolates of slowly growing Mycobacteria(Mycobacterium simiae,Mycobacterium lentflavum and Mycobacterium parascrofulaceu)and rapidly growing Mycobacteria(Mycobacterium chelonae,Mycobacterium mageritense and Mycobacterium vaccae).All the isolates could be identified by multiple homologous sequence.Based on the results of multiple homologous sequence,7 errors were identified by microarray chip,and the identification accuracy of complex groups was 90.0%(153/170).The accuracies of 16S rRNA,rpoB,hsp65 and 3 homologous sequences were 93.5%,98.8%,99.4%and 100.0%,respectively.For 53 isolates of Mycobacterium chelonis/Mycobacterium abscessus complex(MABC)that could not be distinguished by microarray chip,6 isolates of Mycobacterium chelonis,25 isolates of MABC subspecies,20 isolates of MABC Masai subspecies and 2 isolates of MABC bolletii subspecies were identified by 3 homologous sequences.It took 8 h for TAT to be identified by microarray chip and 2 d for all the 170 NTM isolates to be identified.It took 3 d to identify TAT from a single sample by multiple homologous sequence and about 5 d to identify all the 170 NTM isolates.Conclusions Microarray chip could quickly provide accurate results

关 键 词：微阵列芯片法 同源基因序列 非结核分枝杆菌 脓肿分枝杆菌复合群 样本周转时间 

分 类 号：R446.7[医药卫生—诊断学]