交叉引物扩增技术检测痰样本中结核分枝杆菌的应用价值  

Application of cross-priming amplification on direct detection of Mycobacterium tuberculosis DNA from sputum samples

作　　者：王静[1] 刘立宾[1] 岳永宁[1] 胡钰卿[1] 范大鹏[1] 李浩[1] 竺祖军[1] 张艳[1] 夏强[1] 朱敏[1] 

机构地区：[1]杭州市红十字会医院结核病诊疗中心,浙江杭州310003

出　　处：《中国预防医学杂志》2016年第4期265-268,共4页Chinese Preventive Medicine

基　　金：杭州市卫生计生科技计划一般(A类)项目(2015A22);杭州市红十字会医院院内课题资助项目(hhyn201206)

摘　　要：目的采用ROC曲线分析法评估交叉引物扩增技术(CPA)直接检测临床痰样本结核分枝杆菌的检测效能,为临床应用提供参考依据。方法选取肺结核患者225例和非肺结核患者82例,采用CPA进行检测,并同时使用涂片抗酸染色、罗氏培养法、荧光定量PCR技术(FQ-PCR)三种最常用的实验室诊断方法作为对照。根据每种方法对样本检测结果的灵敏度和特异度,以受试者工作曲线(ROC)进行分析比较。结果去除非结核分枝杆菌的26个样本,以临床诊断为金标准,涂片抗酸染色、罗氏培养法、FQPCR技术和CPA的灵敏度分别为46.73%(93/199),63.32%(126/199),69.85%(139/199),70.35%(140/199),假阳性率分别为0%(0/82),0%(0/82),1.22%(1/82),1.22%(1/82)。ROC曲线下的面积分别为0.729、0.817、0.841、0.843,均P<0.01。结论 ROC曲线分析结果显示,交叉引物扩增技术诊断结核病具有较好的准确性,该方法可作为结核病的实验室诊断方法。Objective To assess the application of cross-priming amplification（CPA）for detection of Mycobacterium tuberculosis in sputum samples by using ROC curve analysis. Methods To compare cross-priming amplification（CPA）with acid-fast bacilli（AFB）smear microscopy,Lowenstein-Jensen（L-J）culture and real-time fluorescence quantitative PCR（FQ-PCR）,225 sputum specimens from pulmonary tuberculosis patients and 82 sputum specimens from non-pulmonary tuberculosis patients were collected and tested.ROC analysis was used to evaluate the effectiveness of cross-priming amplification（CPA）. Results Twenty-six samples with nontuberculous mycobacteria were excluded from the study.With clinical diagnosis as the golden standard,the sensitivities of AFS,L-J culture,FQ-PCR and CPA were 46.73%（93/199）,63.32%（126/199）,69.85%（139/199）and 70.35%（140/199）,respectively,and the false positive rates were 0%（0/82）,0%（0/82）,1.22%（1/82）and 1.22%（1/82）,respectively.The areas under ROC curve were 0.729,0.817,0.841 and 0.843. Conclusions CPA is a rapid,sensitive and specific method for detection of Mycobacterium tuberculosis in clinical sputum samples.

关 键 词：结核分枝杆菌 交叉引物扩增技术 痰 ROC曲线 

分 类 号：R378.911[医药卫生—病原生物学]