title: Prevalence and characterization of extended-spectrum beta-lactamases in Klebsiella pneumoniae in Algiers hospitals (Algeria).
abstract: OBJECTIVE
  To determine the prevalence and the diversity of extended-spectrum beta-lactamases (ESBLs) in 196 Klebsiella pneumoniae clinical isolates collected from three hospitals in Algiers.
  
  
  METHODS
  Antibiograms were done on Mueller-Hinton agar plates with the disc-diffusion method and MICs were determined by agar-dilution method. Mating experiments were performed in agar medium. Plasmid DNA was extracted by the alcalin-lysis method. Total DNA was extracted with a Qiagen mini kit and screened for bla(TEM) and bla(CTX-M) genes by PCR. Linkage of bla(CTX-M) genes with insertion sequence ISEcp1B and class 1 integrons was investigated by PCR. PCR products were sequenced by the Sanger method. The epidemiological relationships between ESBL-producing K. pneumoniae isolates were analyzed by ERIC-PCR.
  
  
  RESULTS
  Thirty-nine (19.9%) isolates were found to produce ESBLs belonging to CTX-M-1 group and TEM penicillinases (CTX-M-3, CTX-M-15 and TEM-1). ERIC-PCR analysis showed that the isolates are genetically unrelated. The bla(TEM) and bla(CTX-M) genes as well as aminoglycosides and sulfonamides resistance determinants were found located in self-transferable plasmids of approximately 85 kb. The class 1 integrons and the insertion sequence ISEcp1B were present in the isolates and in their transconjugants. ISEcp1B was found genetically linked to the bla(CTX-M) genes and located 127bp upstream, with the presence of the V and W sequences.
  
  
  CONCLUSIONS
  The study revealed a high rate of ESBL-producing K. pneumoniae in Algerian hospitals, resulting from horizontal dissemination of mobile bla(CTX-M) genes.
authors.affiliation: Laboratoire de biologie cellulaire et moléculaire, faculté des sciences biologiques, université des sciences et de la technologie Houari-Boumédiène, B.P. 32 El-Alia, Bab-Ezzouar 16111, Alger, Algeria.
authors.forename: Y
authors.initials: Y
authors.lastname: Messai
authors.forename: H
authors.initials: H
authors.lastname: Iabadene
authors.forename: T
authors.initials: T
authors.lastname: Benhassine
authors.forename: S
authors.initials: S
authors.lastname: Alouache
authors.forename: M
authors.initials: M
authors.lastname: Tazir
authors.forename: V
authors.initials: V
authors.lastname: Gautier
authors.forename: G
authors.initials: G
authors.lastname: Arlet
authors.forename: R
authors.initials: R
authors.lastname: Bakour
mesh_terms: D000462:Algeria / Q000453:epidemiology; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D003227:Conjugation, Genetic; D003428:Cross Infection / Q000382:microbiology; D024901:Drug Resistance, Multiple, Bacterial / Q000235:genetics; D006801:Humans; D032023:Integrons / Q000235:genetics; D007710:Klebsiella Infections / Q000453:epidemiology / Q000382:microbiology; D007711:Klebsiella pneumoniae / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification; D016133:Polymerase Chain Reaction; D015995:Prevalence; D013379:Substrate Specificity; D018440:beta-Lactam Resistance / Q000235:genetics*; D001618:beta-Lactamases / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D047090:beta-Lactams / Q000494:pharmacology
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D047090:beta-Lactams; D001618:beta-Lactamases