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

• DOI: 10.1007/s101560050045
• Publication Date: 2002-08-25T05:57:49Z
• Authors: Takashi Deguchi; Masayoshi Yamaha; Masahiro Nakano; Mitsuru Yasuda; Yoshinori Nishino; Satoshi Ishihara; Yukimichi Kawada
• Keywords: DNA Topoisomerase IV; 0301 basic medicine; 4-Quinolones; In Vitro Techniques; Polymerase Chain Reaction; 3. Good health; 03 medical and health sciences; Anti-Infective Agents; Drug Resistance, Bacterial; Pseudomonas aeruginosa; Humans; Point Mutation; Genetic Testing; DNA Primers

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.