摘要
目的建立快速检测结核分枝杆菌利福平(RFP)耐药rpoB基因突变的DNA芯片。方法从34株临床痰样本中提取结核分枝杆菌的基因组DNA,采用聚合酶链反应(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特异突变位点的寡核苷酸探针进行杂交,同时与DNA测序法比较。结果用DNA芯片检测出19株耐药株和15株敏感株,且结果与DNA测序相同,准确率可达88.2%,敏感性为78.9%,特异性为100.0%。结论DNA芯片快速检测结核分枝杆菌对RFP的耐药rpoB基因突变是一种操作简便、准确性、敏感性和特异性较高,且易于开展的方法。
Objective To develop a new DNA chip for rapid detection of rpoB mutation rifampin-resistant Mycobacterium tuberculosis isolated from clinical laboratorial specimen. Methods Mycobacterium tuberculosis genome DNA was extracted from 34 sputum samples. The DNA fragment which contained hot mutation sites of rpoB gene was amplified by polymerase chain reaction （PCR） and labeled with cy3 fluorescence. Then it was hybridized with DNA chip. DNA of wild type was used as the control. Results Among 34 clinical sputa specimens, 19 rifampin-resistant strains and 15 rifampin-sensitive strains were found by DNA chip. Comparing with DNA sequencing, the accuracy rate of DNA chip was about 88.2% ; The sensitivity and the specificity was 78.9% and 100.0% respectively. Conclusions DNA chip has high accuracy and sensitivity in detection of rifampin-resistant Mycobacterium tuberculosis and may be applied in clinical diagnosis.
出处
《检验医学》
CAS
北大核心
2008年第4期405-407,共3页
Laboratory Medicine
