MPB64分泌蛋白检测鉴定结核分枝杆菌复合群的应用研究

目的 对应用MPB64分泌蛋白检测鉴定结核分枝杆菌复合群进行方法学评价.方法 对临床分离菌株,取BacT/ALERT 3D系统的液体培养液或改良罗氏固体培养基分离培养菌落室温静置约2 h、浊度为1麦氏单位的生理盐水菌悬液0.1 ml为检测样本,应用胶体金标记抗MPB64单克隆抗体采用免疫层析色谱法进行MPB64分泌蛋白检测,而后与菌种鉴定结果相比较.对参考菌株,则同时采用两种样本进行检测.结果 对23株分枝杆菌参考菌株、267株临床分离菌株进行了检测.①对于参考菌株的两种样本,MPB64检测结果均为:结核分枝杆菌、牛分枝杆菌、田鼠分枝杆菌菌株阳性,其他20株非结核分枝杆菌为阴性.分群鉴定准确度为100%.②在111株BacT/ALERT 3D系统分离培养临床株中,55株为结核分枝杆菌,其中51株MPB64检测阳性;56株为非结核分枝杆菌,其中54株MPB64检测阴性.分群鉴定准确度为94.59%.③在156株改良罗氏培养基分离培养临床菌株中,121株为结核分枝杆菌,其中115株MPB64检测阳性;35株为非结核分枝杆菌,MPB64检测全部阴性.分群鉴定准确度为96.15%.④对于所有研究样本,应用MPB64分泌蛋白检测鉴定结核分枝杆菌复合群的敏感度、特异度与准确度分别为94.41%、98.20%和95.86%.结论 应用MPB64分泌蛋白检测鉴定结核分枝杆菌复合群是一种准确、快速、简便而又不需要任何仪器设备的实用型方法,值得临床推广应用.

Study on application of secretory protein MPB64 for identification of Mycobacterium tuberculosis complex

Objective To evaluate the application of detecting secretory protein MPB64 for the identification of Mycobacterium tuberculosis complex.Methods For clinical isolated strains,0.1 ml of either liquid medium obtained from BacT/ALERT 3D system or bacterial sodium suspension from colonies cultured by L(o)wenstein-Jensen solid medium with 1 Mcfarland standard at room temperature for about two hours was made as samples.Secretory protein MPB64 was detected by immunochromatographic method with colloidal gold labeled anti-MPB64 monoclonal antibody,then the results were compared with the strain identification.For reference strains,both liquid culture medium and bacterial sodium suspension were detected.Results Totally,23 reference strains and 267 clinical isolated strains of mycobacteria were detected.① For the reference strains,Mycobacterium tuberculosis,Mycobacterium bovis and Mycobacterium microti were positive in two kind of samples,and other 20 non tuberculous Mycobacterium were negative.The accuracy of identification was 100 ％.②Among 111 clinical isolated strains from BacT/ALERT 3D system,55 strains were Mycobacterium,in which MPB64 of 51 strains was positive,and 56 strains were non'tuberculous Mycobacterium,in which MPB64 of 54 strains was negative.The accuracy of identification was 94.59 ％.③ Among 156 clinical isolated strains from L(o)wenstein-Jensen solid medium,121 strains were Mycobacterium tuberculosis,in which MPB64 of 115 strains was positive,and 35 strains were non tuberculous Mycobacterium,in which MPB64 was all negative.The accuracy of identification was 96.15 ％.④For all samples,the sensitivity,specificity and accuracy in the identification of Mycobacterium tuberculosis complex by detecting secretory protein MPB64 were 94.41％,98.20％ and 95.86％,respectively.Conclusions Identifying Mycobacterium tuberculosis complex by detecting secretory protein MPB64 was a accurate,fast and simple method.Furthermore,it could be widely used in clinic because of no special instrument.