摘要
目的 开发快速检测耐利福平结核分枝杆菌 (结核菌 )rpoB基因突变的DNA芯片。方法 根据结核菌rpoB基因序列设计探针并制作基因芯片 ,从临床样品中分离出结核菌的基因组DNA ,PCR扩增含有rpoB基因突变位点的特异DNA片段 ,荧光标记后与芯片上含有的检测特异突变位点的寡核苷酸探针进行杂交 ,同时与DNA直接测序法测定序列比较。结果 35株耐利福平结核菌中有 91 4 % (32 / 35 )用直接测序法检测出存在rpoB基因突变 ,DNA芯片的检测效率为 71 4 % (2 5 / 35 )。结论 用DNA芯片检测结核菌对利福平的耐药性具有较高的特异性和敏感性 ,可用于临床结核菌耐药性检测。
Objective To develop a new DNA chip for rapid detection of rpoB gene mutation in rifampin-resistant Mycobacterium tuberculosis isolated from clinical laboratorial specimen. Methods 9 oligonucleotide probes specific for detection of the mutant sequences in rpoB gene were designed. Results Mutations were found in 32 strains (91.4%) of 35 randomly selected rifampin-resistant M. tuberculosis by PCR-direct sequencing technique. 25 mutations (71.4%) were found by DNA chip. Conclusion DNA chip technology has high sensitivity and specificity in detection of rifampin-resistant M. tuberculosis and may be applied in clinical diagnosis.
出处
《中华检验医学杂志》
CAS
CSCD
北大核心
2003年第11期680-682,共3页
Chinese Journal of Laboratory Medicine
基金
香港特区政府创新科技基金 (SERAP :S/P5 99/0 3A)
