利用尿素酶试验测定结核分支杆菌耐药性的方法学研究

Methology to Determinate Drug Resistance of Mycobacterium Tuberculosis by Employing Urease Test

目的:利用尿素酶试验的方法对结核分枝杆菌的耐药性进行检测。方法:采用液体结核分枝杆菌的培养基加尿素和酚红指示剂,高压消毒灭菌,将分离纯培养的结核分枝杆菌接种其中,再加入不同浓度的抗结核分枝杆菌的药物培养,分别设阳性和阴性对照。待阴、阳性对照结果完全清晰时,终止培养,观测结果。结果:结核分枝杆菌、在500cfu/mL以上时,结果在3d～7d内显现;在500条以内时,其结果在培养7d～10d内显现;阳性菌株中,通过加温处死,其尿素酶试验均为阴性;与传统的绝对浓度法比较,差异无显著性(χ2=2.25,P>0.10)。结论:尿素酶试验方法快速、简便,将对结核分枝杆菌耐药性检测提供一种新方法。

Objective：To determinate drug resistance of tubercle bacillus by employing urease test. Methods： The observed purified mycobacterium were inoculated into the specific autoclaved fluid mycobacterium medium with urea and Phenol Red, and incubated with different concentration of anti - tubercle bacillus drugs until the corresponding negative and positive results became apparent. Results： The results became apparent at 3 to 7 days when mount of the incubated mycobacterium is more than 500 cfu/mL, and 7 to 10 days when mount is less than 500 cfu/ml. To avoid the fake positive samples, the positive mycobacteriums were heating - killed and all tested as negative by urease test. Compared with traditional absolute concentration method, urease test method had no statistic differences （Х^2= 2.25, P 〉 0.05）o Conclusion： Urease testing is a convenient and rapid method to inspect drug＇resistance of mycobacterium tuberculosis.