结核分枝杆菌Ag85B mRNA定量检测作为化疗监测指标的初步研究

Measurement of Messenger RNA in Mycobacterium Tuberculosis from Sputum as a Marker for Monitoring the Response to Chemotherapy

目的探讨痰标本中结核分枝杆菌Ag85BmRNA分子作为肺结核患者化疗反应监测指标的可行性和临床价值。方法应用定量逆转录聚合酶链反应（RT—PCR）对15例初治涂阳肺结核患者应用标准短程化疗方案治疗过程中2、4、7、14、30、60d痰标本的结核分枝杆菌Ag85BmRNA进行系列检测，并与培养法（CFU）作比较。结果痰标本中结核分枝杆菌Ag85BmRNA在治疗的前2d下降明显且与CFU的下降基本一致，治疗7dAgS5BmRNA检测阴性11例，治疗14d阴性的12例。治疗30d除1例阳性外，其余朝为阴性，治疗60d时全部阴性。结论结核分枝杆菌Ag85BmRNA在接受治疗的肺结核患者的痰标中快速下降，是活菌数量下降的反映，痰标本中mRNA的变化可作为快速评价化疗反应的分子指标．

Objective To study the clinical value of Ag85B mRNA of M.tuberculosis in the sputa as a marker of monitoring the response of patients with pulmonary tuberculosis to chemotherapy. Method The sputa were collected from 15 smear positive, primary treated patients with pulmonary tuberculosis at 2, 4, 7, 14,30, 60 days after receiving a standard short course ehemotherapie regimen, measured Ag85B mRNA of M. tuberculosis using quantitative reverse-transcribed polymerase chain reaction assay. The results were compared with that using culture method. Results The level of Ag85B mRNA declined rapidly at 2 days after initiation of therapy, which was consistent with viable M. tuberculosis colony counts. Of 15 patients, 11, 12, and 14 had no detectable mRNA at 7, 14, and 30 days after treatment ,respectively. None of sputa had detectable mRNA at 60 days after treatment. Conclusion The rapid disappearance of M. tuberculosis Ag85B mRNA from sputum suggests that it might be a good indicator of microbial viability and a useful marker for rapid evaluation of response to chemotherapy.