用Phage88快速检测结核分枝杆菌

Detection of mycobacterium by using temperate mycobacteriophage

目的 建立一种特异性强、灵敏度高的致病性分枝杆菌的检测及药敏试验方法。方法 应用可表达荧光素酶的结核分枝杆菌噬菌体 Phage88,采用生物发光方法对不同细菌的发光反应和药敏试验进行检测。结果 Phage88特异地对各种分枝杆菌发光 ,对非分枝杆菌发光值很低 ,两者差异有显著性 ,不同的分枝杆菌发光值有差异 :卡介苗的发光值最高 ,结核杆菌的发光值最低 ;在含抗结核药物的培养基中 ,耐药结核杆菌的发光强度比非耐药结核杆菌强 ,其强度有明显差异。结论 用 Phage88噬菌体检测结核分枝杆菌是一种快速、敏感的检测和药敏试验方法。

Objective To set up a fast, sensitive and correct method to detect mycobacterium and drug sensitivity of mycobacterium by using recombinant mycobacteriophages and bioluminescent method. Methods Luciferase activity was detected in different bacteria by using a recombinant temperate mycobacteriophage (Phage88). Then drug susceptibilities of different mycobacterium were assessed by using phage88. Finally the sensitivity and correction rate was compared between luciferase assay and L J medium culture. Results Phage88 has high light production in mycobacterium specifically and only has very low light production in E.coli, and the difference is significant. Among different mycobacterium, BCG has the highest light production and mycobacterium tuberculosis has the lowest light production. The light production of drug resistant mycobacterium strains is much higher than that of drug sensitive strains in culture with antituberculosis drugs, including rifampicin, isoniazid and streptomycin. The duration of drug resistance test is 72 hours, which took much less time than the normal L J method. Conclusions This assay shows a fast, sensitive and correct method to detect mycobacterium strains and their drug sensitivity.