Development of a rapid assay for screening point mutations associated with quinolone resistance in the Pseudomonas aeruginosa parC gene.

Abstract To detect quinolone resistance-associated mutations within the Ser-80 and Glu-84 codons of the Pseudomonas aeruginosa parC gene, we developed a rapid and simple assay based on polymerase chain reaction (PCR) amplification of the region of the parC gene containing the mutation sites and digestion of the PCR products with a restriction enzyme. The mutations generating alterations at Ser-80 and Glu-84 were detected as restriction fragment length polymorphisms of the PCR products digested with Hin fI. Among 22 clinical isolates tested by this assay, mutations at the Ser-80 and Glu-84 codons were detected in all 10 isolates in which the presence of the mutations had been confirmed previously by DNA sequencing. This rapid and simple assay could be a useful screening device for genetic alterations associated with resistance to quinolones in the P. aeruginosa parC gene.