Development of a rapid assay for screening point mutations associated with quinolone resistance in the Pseudomonas aeruginosa parC gene.
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 HinfI. 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.
Deguchi
T
T
Department of Urology, Gifu University School of Medicine, 40 Tsukasa-machi, Gifu 500-8705, Japan. deguchit@cc.gifu-u.ac.jp
Yamaha
M
M
Nakano
M
M
Yasuda
M
M
Nishino
Y
Y
Ishihara
S
S
Kawada
Y
Y
D042462:4-Quinolones; D000890:Anti-Infective Agents / Q000494:pharmacology*; D017931:DNA Primers; D027101:DNA Topoisomerase IV / Q000235:genetics*; D024881:Drug Resistance, Bacterial; D005820:Genetic Testing; D006801:Humans; D066298:In Vitro Techniques; D017354:Point Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D011550:Pseudomonas aeruginosa / Q000187:drug effects* / Q000235:genetics*
D042462:4-Quinolones; D000890:Anti-Infective Agents; D017931:DNA Primers; D027101:DNA Topoisomerase IV