应用DNA芯片法检测耐多药结核分枝杆菌的研究

Rapid Detection of Multidrug-resistant Mycobacterium Tuberculosis by DNA Chip

目的应用DNA芯片技术快速检测耐多药结核菌的常见突变位点,初步评价其临床应用价值。方法以结核分枝杆菌标准株H37RV为对照,应用基因芯片技术、聚合酶链反应-单链构象多态性技术、聚合酶链反应-直接测序技术检测46株耐多药结核分枝杆菌临床分离株katG、rpoB基因。结果 46例耐多药菌株中,应用基因芯片检测出katG基因突变28株,检出率为60.9%(28/46);检测出rpoB基因突变37株,检出率为80.4%(37/46);同时检测出二者突变共25株,检出率为54.3%(25/46)。结论基因芯片技术操作简单、检测快速,对异烟肼、利福平耐药基因检出率较高,可作为临床耐药检测的辅助手段。

Objective To develop and evaluate a new DNA chip diagnosis technique for rapid detection of multidrug-resistant mycobacterium tuberculosis isolated from clinical laboratorial specimen.Methods Compared with the standard strain,46 multidrug-resistant strains were detected by oligonucleotide microchips,PCR-SSCP and analyzed by direct sequencing technique.Results katG mutations were found in 28 strains（60.9%,28/46） detected by DNA chips,rpoB mutations positive in 37 strains（80.4%,37/46）,katG and rpoB simultaneously positive in 25 strains（54.3%,25/46）.Conclusion DNA chip technology has high sensitivity and specificity in detection of multidrug-resistant mycobacterium tuberculosis and may be applied in clinical diagnosis.