NONRADIOACTIVE SINGLE-STRAND CONFORMATION POLYMORPHISM ANALYSIS FOR DETECTION OF FLUOROQUINOLONE RESISTANCE IN MYCOBACTERIA

A simple, rapid, and nonradioactive method for routine detection of fl uoroquinolone resistance in mycobacteria is described. A single-strand conformation polymorphism (SSCP) methodology, based on the use of min i-gels and silver staining of DNA, was optimized for the analysis of d enatured DNA products obtained by polymerase chain reaction (PCR) from the gyrA gene involved in fluoroquinolone resistance in mycobacteria. The method was successfully applied to fluoroquinolone-susceptible an d -resistant laboratory strains of Mycobacterium smegmatis and to clin ical strains of Mycobacterium tuberculosis isolated from patients who developed resistance during the course of fluoroquinolone treatment.