用DNA芯片快速检测结核分支杆菌对利福平的耐药性  

Rapid detection of Mycobacterium tuberculosis resistance to rifampin using DNA chip

作　　者：景奉香[1] 胡忠义[2] 孙悦[1] 孙斌[2] 赵建龙[1] 

机构地区：[1]中国科学院上海冶金研究所,上海200050 [2]上海市结核病防治中心

出　　处：《中华结核和呼吸杂志》2001年第9期551-554,共4页Chinese Journal of Tuberculosis and Respiratory Diseases

基　　金：中国科学院重大项目 (KSCX1 0 6 );上海市科发基金资助项目 ( 99QD14 0 30 )

摘　　要：目的　研制一种新型DNA芯片 ,用于结核分支杆菌对利福平的耐药性快速检测。方法　根据结核分支杆菌rpoB基因的序列信息设计探针并制作DNA芯片 ,PCR扩增并荧光标记包含结核分支杆菌rpoB基因突变热点的目的片段 ,与芯片杂交 ,同时以DNA测序法为对照。结果　 17株随机抽取的耐利福平结核分支杆菌菌株有 14株可用DNA芯片检测出来 ,效率可达 83%;患者痰液中少量的DNA足够进行芯片检测 ,无须进行培养。结论　用DNA芯片来检测结核分支杆菌对利福平的耐药性具有较高的特异性和灵敏度 ,该法快速、精确 。Objective To develop a new method, DNA chip, which can be used for rapid detection of Mycobacterium tuberculosis resistance to RFP. Methods Designing probe according to the sequence of Mycobacterium tuberculosis rpoB gene and fabricating DNA chip The DNA fragment which contains hot mutation sites of rpoB gene was labelled with cy5 fluorescence and amplified by PCR technique Then it was hybridized with DNA chip DNA sequence was used as the control. Results 14 strains were detected out of 17 randomly selected Mycobacterium tuberculosis resistant to RFP using DNA chip The efficiency was 83% A little bit DNA in the sick sputa was sufficient for drug resistance detection without being incubated. Conclusion It showed higher specialty and sensitivity using DNA chip to detect Mycobacterium tuberculosis resistance to RFP This method was rapid and accurate and could be used for clinical detection of RFP resistant strains

关 键 词：结核分支杆菌 RPOB基因 利福平 药物耐受性 DNA芯片 

分 类 号：R446.5[医药卫生—诊断学]