Objective To evaluate the efficacy of the melting curve method in detecting the resistance of Mycobacterium tuberculosis(MTB)to rifampicin and isoniazid,and to analyze the drug resistance characteristics of MTB in Tangshan Area,to provide a basis for clinical treatment of tuberculosis.Methods The resistance of 275 MTB strains of rifampicin and isoniazid was detected by phenotypic antimicrobial susceptibility(microplate method)and molecular antimicrobial susceptibility(melting curve method),respectively,and the results of the two groups were compared and analyzed by the Kappa test.Results The phenotypic resistance rates of rifampicin and isoniazid in Tangshan Area were 14.18％and 28.64％.The rates of molecular resistance were 17.95％and 26.01％.Using the microplate method as the reference method,the sensitivity,specificity,compliance of the melting curve method for the detection of rifampicin resistance were 89.74％,94.02％,and 93.41％,respectively,which were consistent with the microplate method(Kappa=0.754).The sensitivity,specificity,and coincidence rates of isoniazid resistance were 76.32％,93.40％,and 88.64％,respectively,which were of moderate consistency with the microplate method(Kappa=0.656).There was a diversity of rifampicin and isoniazid resistance gene mutations,and the rifampicin resistance gene mutations were mainly codon single region mutations at positions 529-533 of the rpoB gene,with a mutation ratio of 53.06％(26/49).The codon mutation of katG315 was the main mutation of the isoniazid resistance gene,and the mutation proportion was 53.52％(38/71).Conclusion The melting curve method can quickly and accurately detect the resistance of MTB to a variety of drugs,and can be popularized and applied as a reliable method for drug susceptibility detection.
Mycobacterium tuberculosisDrug-resistant tuberculosisMolecular drug resistanceMelting curve methodPCR
NETL
NSTL
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