恒温扩增荧光检测法检测痰标本结核分枝杆菌复合群核酸的应用分析

Application analysis of loop-mediated isothermal amplification fluorescence detection method for detection nucleic acid of Mycobacterium tuberculosis complex in sputum specimens

目的 分析恒温扩增荧光检测法检测痰标本Mtb复合群核酸的应用前景.方法 选取2015年5-8月在四川省三台县疾病预防控制中心门诊就诊的疑似肺结核患者作为研究对象,共计90例.收集研究对象晨痰和随访2个月月末的晨痰标本,共计180份.分别使用恒温扩增荧光检测法、直接痰涂片镜检法、Xpert Mtb/RIF检测法对患者的痰标本进行检测,对检测结果进行比较.同时,对恒温扩增荧光检测法、Xpert Mtb/RIF检测耗时及检测成本进行比较.结果 90例患者中,共获得82例患者的检测结果.痰涂片法、Xpert Mtb/RIF检测法和恒温扩增荧光检测法检测阳性率分别为30.5％（25/82）、41.5％（34/82）、41.5％（34/82）.恒温扩增荧光法与痰涂片法差异有统计学意义（x2 =32.09,P＜0.05）.对恒温扩增荧光检测法与XpertMtb/RIF检测法进行一致性分析,实际一致率为Pv=97.56％,理论一致率Pt=0.51,Kappa值为0.95,具有最强一致性.82例患者痰标本使用恒温扩增荧光检测法,总耗时362 min,检测成本为14 100元;使用Xpert Mtb/RIF检测,总耗时2123min,检测成本31 160元.结论 恒温扩增荧光检测法阳性检出率与Xpert Mtb/RIF相当,同痰涂片检测法相比能提高结核病患者检出率,且恒温扩增荧光法检测速度快、成本低,具有良好的应用推广前景.

Objective To analyse the application prospect of the loop-mediated isothermal amplification fluorescence detection technology in sputum. Methods A total of 90 suspected pulmonary tuberculosis outpatients were enrolled from May to August 2015 in Sichuan Province Santai County Center for Disease Control and Prevention. A total of 180 morning sputum samples were collected. To detect these sputum specimens by loop-mediated isothermal amplification fluorescence detection, sputum smear microscopy and Gene Xpert Mtb/RIF method, respectively. At the same time, the time and costs were compared between the loop-mediated isothermal amplification fluorescence detection and Gene Xpert Mtb/RIF. Results The detection results of 82 cases were obtained in 90 patients. Sputum smear microscopy, Gene Xpert Mtb/RIF method, loop-mediared isothermal amplification fluorescence detection method, positive rate was 30.49 % （25/82）, 41.5 % （34/82）, 41.5 % （34/82） respectively. There were statistically significant differences in loop-mediared isothermal amplification fluorescence detection method and sputum smear microscopy （x2 =32.09, P〈0.05）. Kappa consistency test showed the strongest consistency in the loop-mediated isothermal amplification fluorescence detection and Gene Xpert Mtb/RIF, the actual coincidence rate Po =97.56%, the theory coincidence rates Pe =0.51, Kappa value was 0. 95. In 82 specimens, It takes a total 362 rain and costs 14 100 RMB using loop-mediated isothermal amplification fluorescence detection method, and a total 2123 min and 31 160 RMB costs using Xpert Mtb/RIF. Conclusion The loop-mediated isothermal amplification fluorescence detection method has the consonant positive rate with Gene Xpert Mtb/RIF and better than sputum smear microscopy method. The loop-mediated isothermal amplification fluorescence detection method has a good application prospect with less time and low costs.