【摘要】 目的比较不同标记物在显色法芯片检测结核分支杆菌利福平和异烟肼耐药基因中的影响。方法分别采用地高辛、荧光素和生物素标记引物,建立地高辛-抗地高辛、荧光素-抗荧光素和生物素-亲和素酶呈色法芯片检测结核分支杆菌利福平(RFP)和异烟肼(INH)耐药基因方法,同时用3种方法检测46株结核分支杆菌临床分离株、19种非结核分支杆菌标准株、9种非分支杆菌标准株和结核分支杆菌H37Rv标准株,并对3种方法的敏感性和重复性进行比较。结果经3种方法平行检测,46例结核分支杆菌临床分离株结果完全一致;19种非结核分支杆菌标准株、9种非分支杆菌标准株结果均为阴性,结核分支杆菌H37Rv标准株结果均为野生型;地高辛系统和荧光素系统敏感度为2.0×103拷贝/ml,生物素-亲和素系统敏感度为2.0×104拷贝/ml;3种方法均有良好的重复性。结论3种方法均具有良好的重复性和特异性,生物素系统敏感度略低于地高辛系统和荧光素系统。更多还原

【Abstract】 Objective To compare the effects of different marker in the detection of resistance genes of rifampin and isoniazid in Mycobacterium tuberculosis by using gene chip.Methods Respectively use digoxin,fluorescein and biotin-labeled primers to establish the gene detection system of digoxin/anti-digoxin,fluorescein/anti-fluorescein and Biotin/avidin to detect the resistance gene of rifampin (RFP) and isoniazid (INH) in Mycobacterium tuberculosis,and use three kinds of methods to respectively detect 46 clinical isolated strains of Mycobacterium tuberculosis,19 species of standard strains of non-Mycobacterium tuberculosis,9 standard strains of non-Mycobacterium and H37Rv standard strains of Mycobacterium tuberculosis,and compare the sensitivity and repeatability of three methods.Results Detection by three methods,46 clinical isolated cases of Mycobacterium tuberculosis have the same results,19 species of standard strains of non-Mycobacterium tuberculosis and 9 standard strains of non-Mycobacterium were negative,and H37Rv standard strains of Mycobacterium tuberculosis are all the wildness.The sensitivity of the digoxin,fluorescein and biotin system is 2.0×103,2.0×103,2.0×104copies/ml respectively.Three methods have good repeatability.Conclusion Three methods have good repeatability and specificity,Biotin system sensitivity is slightly lower than digoxin system and luciferase system.更多还原