Rapid detection of fluoroquinolone resistance by isothermal chimeric primer-initiated amplification of nucleic acids from clinical isolates of Neisseria gonorrhoeae

J Microbiol Methods. 2006 Jun;65(3):557-61. doi: 10.1016/j.mimet.2005.10.002. Epub 2005 Nov 8.

Abstract

To ensure a complete response to fluoroquinolone therapy against Neisseria gonorrhoeae infections, rapid susceptibility determinations are required. We assessed a new approach, an isothermal chimeric primer-initiated amplification of nucleic acids (ICAN)/hybrid-chromatography method to detect rapidly fluoroquinolone resistance in N. gonorrhoeae. Comparison of the amplification results with fluoroquinolone minimum inhibitory concentrations (MICs), which were determined by an agar dilution method, showed that the new method accurately determined fluoroquinolone resistance in all ciprofloxacin- and/or gatifloxacin-resistant isolates, but agreed with results based on MICs in only 6 of 8 (75.0%) ciprofloxacin-susceptible and 7 of 12 (58.3%) gatifloxacin-susceptible isolates. Our results suggest that this method can rapidly and reliably detect point mutations in the gyrA gene as well as fluoroquinolone resistance in resistant isolates of N. gonorrhoeae.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anti-Infective Agents / pharmacology*
  • Ciprofloxacin / pharmacology
  • DNA Gyrase / genetics
  • DNA Primers
  • Drug Resistance, Bacterial*
  • Fluoroquinolones / pharmacology*
  • Gatifloxacin
  • Humans
  • Microbial Sensitivity Tests / methods
  • Neisseria gonorrhoeae / drug effects*
  • Nucleic Acid Amplification Techniques / methods*
  • Point Mutation
  • Time Factors

Substances

  • Anti-Infective Agents
  • DNA Primers
  • Fluoroquinolones
  • Ciprofloxacin
  • DNA Gyrase
  • Gatifloxacin