Detection of mycobacterium tuberculosis of rifampin resistant gene using PCR-SSCP

Objective: To study the clinical value of detecting drug resistance of mycobacterium tuberculosis to rifampin by detecting rpoB mutation of mycobacterium tuberculosis using polymerase chain reaction and single stranded conformation polymorphism(PCR-SSCP) technique.Methods: Forty eights strains of rpoB of mycobacterium tuberculosis sensitive to rifampin and forty eights clinic isolated strains of rpoB of mycobacterium tuberculosis resistant to drugs were detected using PCR-SSCP technique.Genes of the rpoBs were amplified with PCR firstly,then identifying the products with SSCP and finding whether there was mutation,analyzed which comparing with the results of control that mycobacterium tuberculosis was sensitive to rifampin.Results: Of the spectra of the 48 clinic isolated strains of mycobacterium tuberculosis resistant to drugs,there were 11 strains identify to that of control.Of the spectra of the 37 strains of mycobacterium tuberculosis resistant to drugs,there were 3 strains identify to that of control;the spectra of the other 34 strains were different with that of control in various degrees.Comparing the results of L-J drug sensitivity of the 48 clinic isolated strains with the parallel results of SSCP,the sensitivity and specificity of detecting mycobacterium tuberculosis resistant to rifampin with PCR-SSCP technique were 91.90% and 90.90% respectively.Conclusions: PCR-SSCP technique is accurate and prompting in identifying rpoB showing mutation of mycobacterium tuberculosis;in clinic it can identify the rifampin-resistance strains quickly and thus provide information to physician to chose sensitive drugs reasonably for effective chemotherapy of tuberculosis.