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目的 探讨mRNA作为结核分支杆菌活菌检测标志的可行性。方法 采用定量聚合酶链反应 (PCR)方法测定结核分支杆菌H3 7RV 在利福平、异烟肼、乙胺丁醇、链霉素、氧氟沙星处理后 2 4、 4 8、 72h 85BmRNA表达水平的变化 ,并与活菌计数方法对比。结果 在异烟肼、利福平和链霉素处理 2 4h时 ,结核分支杆菌H3 7RV 活菌数下降明显 ;在异烟肼、链霉素、乙胺丁醇及氧氟沙星各药各浓度处理 72h时 ,活菌数相似 ,但利福平处理后活菌数较它们低。利福平处理 2 4h后 85BmRNA下降到无药对照的 0 0 2 % ,其他药物处理分别下降到无药对照的 1 %～ 1 0 % ;各药处理 72h后85BmRNA均下降到 1 %以下。结论 mRNA表达水平可以在短时间内反映出用药与无药的区别 ,并与菌落形成单位 (cfu)几乎呈平行关系。mRNA是检测和判定结核分支杆菌“死”“活”的分子标志物
Objective To investigate the feasibility of using mRNA as a marker of viable Mycobacterium tuberculosis. Methods 85B mRNA expression of Mycobacterium tuberculosis H3 7RV was determined by quantitative polymerase chain reaction (PCR) at rifampicin, isoniazid, ethambutol, streptomycin and ofloxacin at 24, 48 and 72h Level changes, and with live bacteria count method contrast. Results After isoflurane, rifampicin and streptomycin treatment for 24 h, the viable count of Mycobacterium tuberculosis H3 7RV decreased significantly. At different concentrations of isoniazid, streptomycin, ethambutol and ofloxacin, The number of viable cells was similar at 72h, but the number of viable cells after rifampin treatment was lower than that of them. After 48 hours of treatment with rifampicin, 85BmRNA decreased to 0 0 2% of the no drug control, and the other drug treatment decreased to 1% ~ 10% of the no drug control respectively; Conclusion The mRNA expression level can reflect the difference between medication and drug-free in a short time and almost parallel with the colony-forming unit (cfu). mRNA is a molecular marker for detection and determination of Mycobacterium tuberculosis “dead” “alive”