结核分支杆菌耐药分子机制的检测技术的研究  

STUDY ON MOLECULAR MECHANISMS AND RAPID DETECTION METHODS OF DRUG-RESISTANT MYCOBACTERIUM TUBERCULOSIS

作　　者：张小刚[1] 何秀云[1] 陈红兵[1] 王仲元[1] 张晓娟[2] 李书琳[3] 

机构地区：[1]解放军第309医院结核病研究室,北京100091 [2]沈阳市胸科医院检验科 [3]吉林市结核病防治研究所

出　　处：《中国现代医学杂志》2003年第10期29-30,34,共3页China Journal of Modern Medicine

摘　　要：目的 :探讨结核分支杆菌对异烟肼、利福平耐药的分子机制 ,建立快速检测耐药分支杆菌基因型的分子生物学方法。方法 :采用聚合酶链反应 -单链构象多态性 (PCR -SSCP)同时检测结核分支杆菌敏感株和耐异烟肼、利福平耐药株的KatG基因、rpoB基因突变。结果 :78株结核分支杆菌临床分离株均未发现KatG、rpoB基因缺失。以结核分支杆菌H37RV为对照 ,3 6株药物敏感株的KatG、rpoB基因SSCP图谱均正常。 42株同时耐异烟肼和利福平的多耐药株中 ,KatG和rpoB基因突变率分别为 66.7% (2 8/4 2 ) ,90 .5 % (3 8/4 2 )。结论 :结核分支杆菌耐异烟肼、利福平耐药性的产生是由于KatG基因和rpoB基因突变所致。应用PCR -SSCP技术可同时快速检测结核分支杆菌异烟肼。Objective:To study on the molecular mechanisms of isoniazid and rifampin resistant tuberculosis and explore its rapid detection methods.Methods:Polymerase chain reation-single strand conformation polymorphism(PCR-SSCP) was employed to dectect KatG and rpoB gene mutation in isoniazid and rifampin resistant tuberculosis isolates. Results:Strain H37RV was used as a control,SSCP pattern of KatG,rpoB PCR amplifcation form 36 drug susceptible strains were all the same as control. No KatG and rpoB gene sequence delection was observel. SSCP patterns of KatG gene were different from control in 28 drug resistant clinical isolates. SSCP patterns of rpoB gene different from control in 38 drug resistant clinical isolates. The sensitivity were 66.7%(KatG),90.5%(rpoB),respectively. Conclusions:The results confirm that mutations of KatG and rpoB genes are the most important mechanisns in isoniazid and rifampin resistant tuberculosis, respectively. It is feasible that using PCR-SSCP to detect drug resistance in mycobacterium tuberculosis.

关 键 词：结核分支杆菌耐药机制异烟肼利福平KatG基因rpoB基因 

分 类 号：R378.911[医药卫生—病原生物学]