免疫色谱法抗-MPB64单克隆抗体检测结核分枝杆菌菌群方法学评价

目的 用免疫色谱法抗-MPB64单克隆抗体快速检测和诊断结核分枝杆菌结核菌群的方法学评价。方法 共收集20株临床标本分离菌株、11株参考菌株和1株结核分枝杆菌标准菌株，应用免疫色谱法检测在培养基上生长的细菌，并和传统鉴定方法、实时荧光探针定量PCR法（FQ-PCR）作比较研究。结果 用免疫色谱法检测1株标准菌株为阳性，检测11株参考菌株发现用该法能完全区分结核和非结核分枝杆菌。对20株临床分离的标本用免疫色谱检出11株结核菌菌群，检出率为55％；用传统鉴定方法检出10株，其中未能检出的一株为混合菌感染；用FQ-PCR法检出10株，其中未能检出的一株为牛结核菌。免疫色谱法能检测到的最低菌浓度为10^5CFU／ml。免疫色谱法、FQ-PCR法和传统鉴定方法的平均耗时分别为15min，1～2d和30d。结论 免疫色谱法是一种简便、快速、准确、敏感和特异性鉴别结核和非结核分枝杆菌的方法，适合在临床推广使用。

The evaluation of immunochromatographic assay using anti-MPB64 monoclonal antibodies in identification of Mycobacterium tuberculosis

Objective To evaluate the immunochromatographic assay (MPB64-ICA) using Anti-MPB64 monoclonal antibodies in identification and diagnosis of Mycobacterium tuberculosis.Methods 20 clinical strains, 11 reference strains and one standard strain of Mycobacterium H37RV were collected and identified by the methods of MPB64-ICA, traditional identification, Fluorogenic Probes and the real-Time PCR(FQ-PCR). The results were compared among these four identification methods.Results When identified by MPB64-ICA, the standard strain, H37RV, showed positive and the 11 reference strains could be completely divided as Mycobacterium tuberculosis and non-ruberculosis Mycobacterium Eleven of 20 (55%) clinical isolates were identified as Mycobacterium tuberculosis by MPB64-ICA.Ten of 20 clinical isolates were identified as Mycobacterium tuberculosis by the traditional identification method and the one unable to be identified was infected by complex Mycobacterium. Ten of 20 clinical isolates were identified as Mycobacterium tuberculosis by FQ-PCR, and the one unable to be identified was Mycobacterium bovis. The detection limit of bacteria for MPB64-ICA was 10~5 CFU/m. The average consuming time of MPB64-ICA, FQ-PCR and traditional method were 15 minutes, 1-2 days and 30 days separately.Conclusion MPB64-ICA was a simple, fast and accurate method to identify Mycobacterium tuberculosis and non-ruberculosis. It could be used in clinic widely.