Value of two molecular methods for detecting resistance of Mycobacterium tuberculosis to rifampicin and isoniazid
Objective To analyze the drug resistance of Mycobacterium tuberculosis(MTB)infected patients in Nanjing,and evaluate the clinical value of fluorescence PCR melting curve method and PCR linear hybridization enzyme chromogenesis assay in detecting the resistance of Mycobacterium tuberculosis to rifampicin(RFP)and isoniazid(INH).Methods A total of 151 clinical isolates of Mycobacterium tuberculosis were collected in the Nanjing Public Health Medical Center.The drug resistance of MTB to RFP and INH were detected by absolute concentration drug sensitivity test,fluorescence PCR melting curve method and PCR linear hybridization enzyme chromogenesis assay.The diagnostic efficacy of the above two molecular biology methods was evaluated using the results of phenotypic susceptibility test.Results Among the 151 clinical isolates of MTB,phenotypic resistance results showed that 6 were RFP single drug resistant,9 were INH single drug resistant,and 43 were multidrug resistant.Using absolute concentration drug sensitivity test as standard,the sensitivity,specificity,coincidence rate and κ value of MTB to RFP and INH resistance detected by fluorescence PCR melting curve method were 100.00％and 94.23％,96.08％and 96.97％,97.35％and 96.02％,0.94 and 0.91,respectively.Using absolute concentration drug sensitivity test as standard,the sensitivity,specificity,coincidence rate and K value of MTB to RFP and INH resistance detected by PCR linear hybridization enzyme chromogenesis assay were 89.79％and 86.54％,96.08％and 94.12％,94.04％and 93.38％,0.86 and 0.85,respectively.There were no significant differences between the two methods(x2=0.182,0.114;all P＞0.05).Conclusions The fluorescent PCR melting curve method and the PCR-linear hybridization enzyme colorimetric method had the same diagnostic efficacy in detecting rifampicin and isoniazid resistance,had good consistency with the phenotypic drug sensitivity test,and could make up for the limitations of the phenotypic drug sensitivity test in detecting mutations of low-resistance strains.They could be used for early and rapid diagnosis of tuberculosis and provide a basis for timely and accurate clinical anti-tuberculosis treatment.
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