mRNA定量法检测结核分枝杆菌药物敏感性的应用评价  

Clinical evaluation of quantitative analysis of 85B mRNA in drug susceptibility testing of M.tuberculosis

作　　者：蔡龙[1] 施华萍[1] 马纪林[1] 程建玲[1] 张艳[1] 

机构地区：[1]浙江省杭州市红十字会医院,310000

出　　处：《实验与检验医学》2008年第1期36-38,78,共4页Experimental and Laboratory Medicine

基　　金：浙江省医药卫生科技计划项目资助(基金编号:2004B171)

摘　　要：目的评价应用mRNA定量法进行结核分枝杆菌利福平、异烟肼药物敏感性分析的临床应用价值。方法应用定量聚合酶链反应(PCR)方法检测三株结核分枝杆菌标准株在利福平、异烟肼处理24小时后85B mRNA表达水平的变化;并以不同的结果判断标准,比较本方法和绝对浓度法对87株临床分离菌株利福平、异烟肼药敏试验检测结果的差异性。结果利福平或异烟肼敏感株在相应的药物处理24小时后85B mRNA表达水平明显下降,分别为无药对照的0.01%和0.35%以下,而耐药株无明显变化。以85B mRNA的表达水平下降到无药对照的1%以下为敏感,大于10%为耐药,1%～10%之间为可疑进行判断,本方法与绝对浓度法有一致的检测结果。结论85B mRNA可以作为结核分枝杆菌药物敏感性试验的分子标志,应用85B mRNA检测结核分枝杆菌的耐药性是一种快速有效的检测方法。Objective To evaluate the clinical value of quantitative analysis of 85B mRNA in drug susceptibility testing of M.tuberculosis （MTB）. Methods The levels of 85B mRNA was observed at 24 hours after three standard strains of MTB treated with either no drug, 1.0μg/ml isoniazid（INH）, or 4μg/ml rifampin（RlF）. With different judgment standards, the detection results of RIF, INH susceptibility with the methods of RT-PCR and absolute concentration were analyzed in 87 clinical isolates. Results After exposure of sensitive strains to RIF or INH for 24 hours, the levels of 85B mRNA reduced respectively to 〈0.01% and 〈 0.35%, compared with those present in control cultures without drug. In contrast, the resistant strains demonstrated no reduction over the same period. According to the standard, it was sensitive when the level of 85B mRNA was less than 1% of that on the control culture, resistant as over 10%, and uncertainty as 1%-10%, the method produced consistency results with absolution concentration. Conclusion The quantitative analysis of 85B mRNA by reverse transcription-PCR is a rapid and effective method in susceptibility test of MTB.

关 键 词：结核分枝杆菌 信使核糖核酸 逆转录聚合酶链反应 药敏试验 

分 类 号：R446.5[医药卫生—诊断学] Q522.2[医药卫生—临床医学]