三种检测技术鉴别结核分枝杆菌复合群与非结核分枝杆菌的效能评价  

Application evaluation of three methods for identification between Mycobacterium tuberculosis complex and nontuberculous mycobacteria

作　　者：易俊莉[1] 杨新宇[1] 张洁[1] 田丽丽[1] 丁北川[1] 武文清[1] YI Jun-li;YANG Xin-yu;ZHANG Jie;TIAN Li-li;DING Bei-chuan;WU Wen-qing(Beijing Research Institute for Tuberculosis Control,Beijing 100035,China)

机构地区：[1]北京结核病控制研究所,100035

出　　处：《结核与肺部疾病杂志》2020年第4期240-244,共5页Journal of Tuberculosis and Lung Disease

基　　金：首都卫生发展科研专项(2018-1-1041)。

摘　　要：目的评价对硝基苯甲酸/噻吩-2-羧酸肼(PNB/TCH)生长试验法、结核分枝杆菌抗原(MPB64)检测法、PCR-荧光探针法在结核分枝杆菌复合群(MTBC)与非结核分枝杆菌(NTM)鉴别中的应用价值。方法11株标准菌株包括MTB标准株H37Rv及10株NTM标准菌株均来源于国家结核病参比实验室。238株临床分离株为北京结核病控制研究所2019年1—12月门诊患者培养阳性冻存菌株,经涂片抗酸染色均为阳性。应用PNB/TCH生长试验法、MPB64检测法、PCR-荧光探针法及微阵列基因芯片法对11株标准菌株及238株临床分离株进行鉴定;以微阵列基因芯片法菌种鉴定结果为参照,评价PNB/TCH生长试验法、MPB64检测法、PCR-荧光探针法鉴别MTBC与NTM的效能。结果以微阵列基因芯片法菌种鉴定结果为参照,PNB/TCH生长试验法、MPB64检测法、PCR-荧光探针法检测MTBC的敏感度分别为100.0%(206/206)、98.5%(203/206)、100.0%(206/206);特异度分别为96.9%(31/32)、100.0%(32/32)、100.0%(32/32);符合率分别为99.6%(237/238)、98.7%(235/238)、100.0%(238/238);Kappa值分别为0.98、0.95、1.00。PNB/TCH生长试验法、MPB64检测法、PCR-荧光探针法检测周期分别为28d、0.5h、0.5d,检测平均成本分别为20、40、60元。PNB/TCH生长试验法和MPB64检测法需要得到培养物才能进行检测,而PCR-荧光探针法需要实验室及操作人员具有专业的核酸检测能力。结论PNB/TCH生长试验法、MPB64检测法、PCR-荧光探针法均可用于MTBC与NTM的鉴别。PCR-荧光探针法敏感度、特异度较高,检测结果更为准确可靠。MPB64检测法操作简便、快速、价格低廉,适合基层实验室进行初步鉴定使用。Objective To evaluate the application of PNB/TCH growth test,Mycobacterium tuberculosis antigen(MPB64)detection and PCR-fluorescent probe method in identification of Mycobacterium tuberculosis complex(MTBC)and non-tuberculous mycobacteria(NTM).Methods A total of 11 standard strains including MTB H37 Rv and 10 NTM standard strains were all from the National Tuberculosis Reference Laboratory.And 238 clinical isolates were frozen positive strains cultured from outpatients of Beijing Research Institute for Tuberculosis Control from January to December 2019,all of them were positive in acid fast smear staining.Eleven standard strains and 238 clinical isolates were identified by PNB/TCH growth test,MPB64 detection,PCR-fluorescent probe method and gene chip technology.Based on the identification results of microarray gene chip method,the efficiency of PNB/TCH growth test,MPB64 detection method and PCR-fluorescent probe method to identify MTBC and NTM was evaluated.Results Compared to strain identification with gene chip technology,the sensitivities of PNB/TCH growth test,MPB64 detection and PCR-fluorescent probe method were 100.0%(206/206),98.5%(203/206)and 100.0%(206/206),respectively;the specificities were 96.9%(31/32),100.0%(32/32),100.0%(32/32).The coincidence rates were 99.6%(237/238),98.7%(235/238)and 100.0%(238/238),respectively;the Kappavalues were 0.98,0.95 and 1.00,respectively.The detection times of PNB/TCH growth test,MPB64 detection method and PCR-fluorescent probe method were 28 days,half an hour,and half of the day,respectively;the average detection costs were RMB 20 yuan,40 yuan and 60 yuan.Both PNB/TCH growth test method and MPB64 detection method need cultures,while PCR-fluorescent probe method requires laboratories and operators with professional ability of nucleic acid detection.Conclusion PNB/TCH growth test,MPB64 detection and PCR-fluorescent probe method could be used to identify MTBC and NTM.PCR-fluorescent probe method has high sensitivity and specificity,and the detection results are more accur

关 键 词：分枝杆菌 结核 分枝杆菌 非典型性 诊断 鉴别 实验室技术和方法 评价研究 

分 类 号：R446.5[医药卫生—诊断学]