MALDI-TOF MS在NTM临床分离株快速鉴定中的应用研究  

Application of MALDI-TOF MS in rapid identification of NTM clinical isolates

作　　者：黄琨波 陈桂春 林玉玲 陈清清[1] 李科 张建明 HUANG Kunbo;CHEN Guichun;LIN Yuling;CHEN Qingqing;LI Ke;ZHANG Jianming(Department of Laboratory,Quanzhou First Hospital Affiliated to Fujian Medical University,Quanzhou,Fujian 362000,China;Department of Cardiology,Quanzhou First Hospital Affiliated to Fujian Medical University,Quanzhou,Fujian 362000,China)

机构地区：[1]福建医科大学附属泉州第一医院检验科,福建泉州362000 [2]福建医科大学附属泉州第一医院心内科,福建泉州362000

出　　处：《检验医学与临床》2023年第15期2162-2165,2169,共5页Laboratory Medicine and Clinic

基　　金：福建省卫生健康中青年骨干人才培养项目(2020GGA075);福建省泉州市科技计划项目(2021N059S)。

摘　　要：目的 建立临床非结核分枝杆菌(NTM)分离株的快速鉴定方法,为临床NTM病的诊断和治疗提供参考。方法 收集2018-2020年泉州某三甲医院的临床疑似NTM感染患者的痰液、肺泡灌洗液、胸部穿刺液和脑脊液等标本,借助基质辅助激光解析/电离飞行时间质谱(MALDI-TOF MS)技术,通过改良前处理方法(甲酸提取法),摸索上机检测条件,建立临床NTM分离株的快速鉴定方法,并以16S rRNA测序为金标准,验证鉴定的准确性。结果 收集的50株临床NTM菌株,采用传统(常规)处理方法进行质谱鉴定,仅鉴定出4株,检出率为8.0%,而采用甲酸提取法共鉴定出24株,检出率为48.0%,两种方法检出率比较,差异有统计学意义(P<0.01),且改良处理后鉴定出的24株鉴定结果与PCR测序结果完全一致,符合率为100.0%。结论 初步建立的NTM甲酸提取前处理的质谱鉴定方法检出率明显提高,具有一定的临床应用价值。Objective To establish a rapid identification method for clinical non-tuberculous mycobacterium(NTM)isolates and provide reference for the diagnosis and treatment of clinical NTM disease.Methods Samples such as sputum,alveolar lavage fluid,chest puncture fluid,and cerebrospinal fluid were collected from patients with clinically suspected NTM infection in 2018-2020 in a tertiary hospital of Quanzhou,China.With the help of Matrix-Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry(MALDI-TOF-MS)technology,a rapid identification method of clinical NTM isolates was established by improving the pretreatment method(the improved formic acid extraction method)and exploring the detection conditions.The accuracy of identification was verified by 16S rRNA sequencing as the gold standard.Results Among the 50 clinical NTM strains collected,only 4 strains were identified by mass spectrometry using the traditional(coventional)treatment method with detection rate of 8.0%,while 24 strains were identified by mass spectrometry using the improved formic acid extraction method after pretreatment with detection rate of 48.0%.The difference of detection rate between the two methods was statistically significant(P<0.01),and the identification results of the 24 strains identified by the improved treatment were completely consistent with the sequencing results of PCR,with compliance rate of 100.0%.Conclusion The detection rate of the mass spectrometric identification method of NTM formic acid pretreated substance is significantly improved,which has certain clinical application value.

关 键 词：基质辅助激光解析/电离飞行时间质谱 非结核分枝杆菌 鉴定 

分 类 号：R446.5[医药卫生—诊断学]