GENETIC MUTATION PROFILE OF gyrA AND parC GENE IN CIPROFLOXACIN-RESISTANCE Pseudomonas aeruginosa

Pseudomonas aeruginosa is a gram-negative bacterium that continues to be a major cause of opportunistic nosocomial infections, causing around 9–10% of infections in hospital. Nosocomial infections caused by Pseudomonas aeruginosa are frequently life threatening and often challenging to treat. A major reason for its prominence as a pathogen is its high intrinsic resistance to antibiotics. Quinolone resistance is increasing with reported frequencies of 12–20%. In most reported there was a missense mutation in the quinolone target (the gyrA subunit of DNA gyrase) at codon 83 (T83I), although other mutations are sometimes observed. Higher levels of resistance may involve additional mutations in gyrB (DNA gyrase B subunit) or parC (topoisomerase IV). Purpose of this study is to determining the genetic mutation of gyrA gene region and parC in Pseudomonas aeruginosa isolates that resistant to ciprofloxacin. Sensitivity testing of bacteria used broth dilution method (BD Phoenix) and Kirby-Bauer methods; DNA extraction used Extract N-AmpTM Blood PCR Kit; QRDR gyrA and parC amplifications used primer form 1st Base OligonucleotideTM with thermal cycler BioRadicycler; agarose gel elektrophoresis; DNA labelling used QiaQuick PCR Column (QIAGEN, Valecia, CA); DNA sequences used Genetic Analyzer ABI Prisma 310, sequencing result is processed used Genetic program version 9.0 and the result were compared with control. Sequencing results predicted changes of an amino acid in the QRDR of DNA gyrase (gyrA) and topoisomerase IV (parC), all isolates possessed mulitple mutation in gyrA and parC. gyrA gene; Glu-53 →Ser, Thr-83 →Ile, Gly-105 →Ala, Glu-106 →Arg, Ala-136 →Gly, parC gene; Asp-35 →Gly, Asp-62 →Asn, Ser-65 →Ala, Ser-87 →Leu, Ser87 →Trp, and additional silent mutation in all isolates.(FMI 2014;50 1-5)