Prevalence and characterization of extended-spectrum beta-lactamases in Klebsiella pneumoniae in Algiers hospitals (Algeria).
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.
Messai
Y
Y
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.
Iabadene
H
H
Benhassine
T
T
Alouache
S
S
Tazir
M
M
Gautier
V
V
Arlet
G
G
Bakour
R
R
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
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D047090:beta-Lactams; D001618:beta-Lactamases