Lü B, Fu Z, Xu S
Institute of Environmental Medicine, Tongji Medical University, Wuhan 430030, China.
Zhonghua Jie He He Hu Xi Za Zhi. 2000 Aug;23(8):480-4.
To set up a fast, sensitive and specific way to detect mycobacteria and rifampicin susceptibility to mycobacteria by using recombinant mycobacteriophages and bioluminescent method.
Firstly detected the luciferase activity in different bacteria by using mycobacteriophages, then assessed the best drug concentration of rifampicin for drug susceptibility test in luciferase reporter assay, and lastly used the above conditions to decide rifampicin susceptibilities to different mycobacteria and compare the result with L-J medium culture.
Bacteriophages had high light production specifically in mycobacteria and only very low light production in E. coli, the difference being obvious. The light production of rifampicin-resisitant mycobacterium strains was much higher than that of sensitive strains in culture with rifampicin(P < 0.05). Different drug concentrations of rifampicin were used to optimize drug concentration for rifampicin drug susceptibility test and the optimum concentration was found to be 2 micrograms/ml. The correlation of drug susceptibility test between luciferase reporter phages to L-J medium in standard strains and clinical isolates was the sarce. Temperature sensitive Phage 88 was more sensitive than Phage 40(P < 0.05). The time for drug resistance test was 72 hours.
Luciferase reporter phage can detect mycobacteria specifically and can be used in rifampicin susceptibility test. This assay is a fast, sensitive and specific method to detect mycobacterium strains and their resistance to rifampicin.
利用重组分枝杆菌噬菌体和生物发光法建立一种快速、灵敏、特异的检测分枝杆菌及分枝杆菌对利福平敏感性的方法。
首先用分枝杆菌噬菌体检测不同细菌中的荧光素酶活性，然后在荧光素酶报告基因检测中评估利福平药敏试验的最佳药物浓度，最后用上述条件判定不同分枝杆菌对利福平的敏感性，并与罗氏培养基培养结果进行比较。
噬菌体在分枝杆菌中产生高亮度发光，在大肠杆菌中产生的光非常低，差异明显。在含利福平培养中，耐利福平分枝杆菌菌株的发光量远高于敏感菌株（P<0.05）。采用不同药物浓度的利福平优化利福平药敏试验的药物浓度，发现最佳浓度为2微克/毫升。荧光素酶报告噬菌体与罗氏培养基对标准菌株和临床分离株的药敏试验相关性良好。温度敏感噬菌体88比噬菌体40更敏感（P<0.05）。耐药试验时间为72小时。
荧光素酶报告噬菌体可特异性检测分枝杆菌，可用于利福平药敏试验。该检测方法是一种快速、灵敏、特异的检测分枝杆菌菌株及其对利福平耐药性的方法。
