title: Molecular characterization of two high-level ceftriaxone-resistant Neisseria gonorrhoeae isolates detected in Catalonia, Spain.
abstract: OBJECTIVE
  The aim of this study was to characterize the first two extended-spectrum cephalosporin-resistant and multidrug-resistant (MDR) Neisseria gonorrhoeae isolates collected from two sexually related patients (men who have sex with men) in Spain.
  
  
  METHODS
  Antimicrobial susceptibility was studied by Etest. Genes involved in quinolone, ceftriaxone and multidrug resistance were amplified by PCR and sequenced in both directions. The isolates were typed by N. gonorrhoeae multi-antigen sequence typing (NG-MAST).
  
  
  RESULTS
  The two isolates had the same MDR profile, showing resistance to penicillin (MIC 0.094 mg/L; β-lactamase negative), ceftriaxone (MIC 1.5 mg/L), cefixime (MIC 1.5 mg/L), cefotaxime (MIC 1 mg/L), ciprofloxacin (MIC >32 mg/L) and tetracycline (MIC 1.5 mg/L). NG-MAST showed that both isolates belonged to sequence type (ST) 1407 (porB-908 and tbpB-110). Ciprofloxacin resistance was due to amino acid substitutions in GyrA (S91F and D95G) and ParC (S87R). An A deletion in the promoter of the MtrCDE efflux pump (mtrR) was detected. No changes were detected in the pilQ gene. The outer membrane protein PorB showed two substitutions at G120K and A121N. An L421P substitution was observed in the PBP1A (ponA) sequence. The sequence of PBP2 (penA) showed a mosaic structure related to genotype XXXIV with a single additional amino acid substitution (A501P). This genotype was identical to a recently described French isolate (F89).
  
  
  CONCLUSIONS
  This is the first reported case of high-level extended-spectrum cephalosporin-resistant N. gonorrhoeae transmission. The molecular typing and MDR genotype suggest possible European spread of this strain, highlighting the need for surveillance and the importance of testing the susceptibility of N. gonorrhoeae to extended-spectrum cephalosporins.
authors.affiliation: Microbiology Department,  Hospital Universitari de Bellvitge-Universitat de Barcelona-IDIBELL, L'Hospitalet de Llobregat, Barcelona,  Spain.
authors.forename: Jordi
authors.initials: J
authors.lastname: Cámara
authors.forename: Judit
authors.initials: J
authors.lastname: Serra
authors.forename: Josefina
authors.initials: J
authors.lastname: Ayats
authors.forename: Teresa
authors.initials: T
authors.lastname: Bastida
authors.forename: Dolors
authors.initials: D
authors.lastname: Carnicer-Pont
authors.forename: Antònia
authors.initials: A
authors.lastname: Andreu
authors.forename: Carmen
authors.initials: C
authors.lastname: Ardanuy
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D002443:Ceftriaxone / Q000494:pharmacology*; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D005838:Genotype; D006069:Gonorrhea / Q000453:epidemiology* / Q000382:microbiology; D018451:Homosexuality, Male; D006801:Humans; D008297:Male; D008826:Microbial Sensitivity Tests; D017720:Molecular Epidemiology; D058889:Molecular Typing; D009344:Neisseria gonorrhoeae / Q000145:classification* / Q000187:drug effects / Q000235:genetics* / Q000302:isolation & purification; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D013030:Spain / Q000453:epidemiology; D018440:beta-Lactam Resistance
chemical_list: D000900:Anti-Bacterial Agents; D004269:DNA, Bacterial; D002443:Ceftriaxone