Multiple Real-time PCR Detection of 3 Drug Resistance Mutations in Mycobacterium Tuberculosis with Displacing Probes

[Objective] To establish PCR assay and evaluate the feasibility of displacing probes and detect related resistance mutations in DNA extracts from sputum samples.[Method] Displacing probes,real-time PCR and multiplex PCR technology were used and 3 genes(rpsL,katG and embB) associated with streptomycin,isoniazid and ethambutol resistance were detected respectively.[Results] Each test of 5 copies mycobacterium tuberculosis can be detected and 9 mycobacterium tuberculosis isolates and 118 sputum samples were evaluated with this assay.All results of real-time PCR assay were consistent with the ARMS assay.[Conclusions] Comparing with classical methods,this assay is of rapid,high sensitivity and specificity.It can be used for high-throughout screening for the mutations of drug resistance in mycobacterium tuberculosis.