PCR实验联合MPB64免疫胶体金检测在分枝杆菌菌种鉴定中的应用

Application of PCR experiment combined with MPB64 immunocolloidal gold detection in identification of mycobacterial strains

目的评价PCR荧光定量实验联合MPB64免疫胶体金检测在分枝杆菌菌种鉴定中的应用价值。方法以分枝杆菌罗氏培养基TCH/PNB生长实验为“金标准”,选取287例河南省胸科医院2018年6月~12月结核分枝杆菌培养菌株,进行PCR和MPB64免疫胶体金检测,分析两种菌种鉴定能力。结果胶体金法、PCR荧光定量法与TCH/PNB实验结果一致率分别是88.9%和99.3%,胶体金法鉴定结核分枝杆菌敏感度、特异度、阳性预测值和阴性预测值分别是94.0%、100.0%、93.7%、68.6%。延长培养后复查MPB64的阴性预测值是87.5%,联合PCR荧光定量实验所得结果与“金标准”符合率100.0%。结论MPB64免疫胶体金法在分枝杆菌菌种鉴定中假阴性率较高,可选择重复实验或者联合PCR实验进行菌种鉴定,方法简便、快速、价格低廉,可推广基层医院应用。

Objective Toevaluate the application of PCR with MPB64 antigen detection of immune colloidal gold method foridentification of Mycobacterium.Methods Taking the Lwenstein-Jensen medium and PNB/TCH growth test as the gold standard,the 287 clinical isolated mycobacterium tuberculosis from June to December in Henan provincial chest hospital were tested by PCR,andwas compared with the method of MPB64 antigen detection.Results Compared with TCH/PNB identification test,the coincidence rate of immune colloidal gold method and sequencing method was 88.9%and 99.3%.The sensitivity,specificity,positive predictive value(PPV)and negative predictive value(NPV)of immune colloidal gold methodidentification were 94.0%、100.0%、93.7%、68.6%.The negative predictive value of MPB64 after prolonged culture was 87.5%,and the coincidence rate between the resultsof combined PCRfluorescence quantitative experiment and"gold standard"was 100.0%.Conclusions The MPB64 immunocolloidal gold method has higher false negative rate in identification of Mycobacterium,so repeated experiment or combined PCR experiment can be selected.The method is simple,rapid and cheap and could be applied in identification of Mycobacterium.