摘要
目的 评价应用mRNA定量法进行结核分枝杆菌利福平敏感性分析的可行性和应用价值。方法 定量检测结核分枝杆菌85B mRNA,分别以mRNA拷贝数百分比(使用利福平药物后mRNA拷贝数占用药前的百分比)为1％和10％作为耐药临界指标分析53株结核分枝杆菌临床分离株对利福平的敏感性,并与绝对浓度法作比较,其中29株为利福平耐药株、24株为利福平敏感株。结果 53株结核分枝杆菌临床分离株mRNA定量法检测:在利福平浓度为1μg/ml的条件下分别以1％和10％为耐药临界比例时,与绝对浓度法相比准确性分别为70％、81％,敏感性分别为:100％、100％,特异性分别为33％、58％;在利福平浓度为2μg/ml的条件下分别以1％和10％为耐药临界比例时,与绝对浓度法相比准确性分别为85%、93％,敏感性分别为100％、97％,特异性分别为67％、88％,在利福平浓度为4μg/ml的条件下分别以1％和10％为耐药临界比例时在与绝对浓度法相比准确性分别为93%、93％,敏感性分别为93％、93％,特异性分别为92％、92％。结论 mRNA定量法用于结核分枝杆菌药物敏感性分析具有快速、可定量等优点,建议mRNA定量法测定利福平敏感性采用临界药物浓度为2 μg/ml、耐药临界比例为10％。
Objective To evaluate the significance of quantitative analysis of Mycobacterium. tuberculosis mRNA in the test of susceptibility of Mycobacterium tuberculosis strains to rifampin. Methods The susceptibility to rifampin of fifty-three clinical isolated strains was test by the percentage of Mycobacterium tuberculosis 85B mRNA copies before and after the use of rifampin, and 1% and 10% were used as the standards for drug resistance, which was compared with the absolute concentration method. Among them, 29 were rifampin resistant strains and 24 rifampin sensitive strains. Results When the concentration of rifampin was 1 μg/ml,and 1% and 10% were used as the standards, the accuracy of quantitative analysis of Mycobacterium tuberculosis mRNA with the absolute concentration method was 70% and 81 % respectively, and the sensitivity of quantitative analysis of Mycobacterium tuberculosis mRNA was 100% and 100% respectively, while the specificity of quantitative analysis of Mycobacterium tuberculosis mRNA was 33% and 58%. When the concentration of rifampin was 2 μg/ml, the accuracy of quantitative analysis of mRNA with the absolute concentration method was 85% and 93% respectively, and the sensitivity was 100% and 97% respectively, while the specificity was 67% and 88% respectively. When the concentration of rifampin was 4 μg/ml, the accuracy was 93% and 93% ,and the sensitivity was 93% and 93% , while the specificity was 92% and 92%. Conclusions Quantitative analysis of Mycobacterium tuberculosis mRNA was a rapid, sensitive method in rifampin resistance screening. We suggest that the critical concentration of rifampin be 2 μg/ml, and the critical proportion of mRNA copy be 10%.
出处
《中华结核和呼吸杂志》
CAS
CSCD
北大核心
2004年第8期549-552,共4页
Chinese Journal of Tuberculosis and Respiratory Diseases
关键词
mRNA定量法
结核分枝杆菌
利福平
敏感性
Mycobacterium tuberculosis
RNA, messenger
Reverse transcriptase polymerase chain reaction
Rifampin
Drug to lerance
