title: DNA gyrase gene in Neisseria gonorrhoeae as indicator for resistance to ciprofloxacin and species verification.
abstract: We have identified a unique region of eight amino acids in the quinolone resistance-determining region in the gyrA gene in Neisseria gonorrhoeae as an indicator of resistance to fluoroquinolones. We sequenced that region by the Pyrosequencing technology in 46 N. gonorrhoeae strains and 11 urine samples positive in AMPLICOR N. gonorrhoeae polymerase chain reaction (Roche Diagnostics), with corresponding isolates of N. gonorrhoeae. The results showed that 28 samples with minimum inhibitory concentration (MIC) of ciprofloxacin >1 mg/L had mutations in positions 91 and 95. Fifteen samples with MIC 0.125-1.0 mg/L had either one or both of the mutations. The 14 susceptible samples had no mutations. The target region also discriminates N. gonorrhoeae from other species of Neisseria. Our conclusion is that gyrA is an indicator of resistance to ciprofloxacin in N. gonorrhoeae and sequencing by Pyrosequencing technology is a suitable tool for analysis of DNA in urine samples.
authors.affiliation: Department of Clinical Bacteriology, Division of Laboratory Medicine, F72 Huddinge University Hospital, SE-141 86 Stockholm, Sweden. emma.lindback@karolinska.se
authors.forename: Emma
authors.initials: E
authors.lastname: Lindbäck
authors.forename: Baback
authors.initials: B
authors.lastname: Gharizadeh
authors.forename: Ferda
authors.initials: F
authors.lastname: Ataker
authors.forename: Asa
authors.initials: A
authors.lastname: Airell
authors.forename: Shah
authors.initials: S
authors.lastname: Jalal
authors.forename: Pål
authors.initials: P
authors.lastname: Nyrén
authors.forename: Bengt
authors.initials: B
authors.lastname: Wretlind
mesh_terms: D002939:Ciprofloxacin / Q000494:pharmacology*; D027081:DNA Gyrase / Q000235:genetics*; D027101:DNA Topoisomerase IV / Q000235:genetics; D024881:Drug Resistance, Bacterial / Q000235:genetics; D009344:Neisseria gonorrhoeae / Q000187:drug effects* / Q000235:genetics*; D017422:Sequence Analysis, DNA
chemical_list: D002939:Ciprofloxacin; D027101:DNA Topoisomerase IV; D027081:DNA Gyrase