title: First report of nosocomial infection caused by Klebsiella pneumoniae ST147 producing OXA-48 and VEB-8 β-lactamases in Tunisia.
abstract: The aim of this study was to determine the origin of virulence and multiresistance of a Klebsiella pneumoniae isolate from an abdominal wound infection of a patient with a gunshot injury in the thoracoabdominal region. The isolate was identified using biochemical tests and Phoenix™ automated system and was confirmed by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF/MS). MICs of each antibiotic were determined by Etest. Screening for carbapenemase production was performed by the modified Hodge test and was confirmed by PCR amplification. Virulence factors were also studied. Plasmid replicon typing was used to classify Incompatibility (Inc) plasmids harbouring the resistance genes. The transferability of each plasmid was determined by conjugation using Escherichia coli J53. Finally, multilocus sequence typing (MLST) was performed to determine the ST of the strain. The bacterial isolate was identified as K. pneumoniae and was named KPM2, carrying entB, ybtS, mrkD and ycfM virulence genes, but it did not overexpress OqxAB. Isolate KPM2 belonged to ST147 and was classified as resistant to all of the tested antibiotics with MICs above the clinical breakpoints. These resistances were due to production of OXA-48, CMY-2, TEM-1, CTX-M-15 and VEB-8 β-lactamases. Genetic and molecular studies showed that blaOXA-48 was embedded in transposon Tn1999.2 and was carried by a conjugative IncL/M plasmid of ca. 60kb; blaVEB-8 was harboured on a conjugative IncA/C plasmid of ca. 120kb. This study confirmed that the resistance conferred by OXA-48 and VEB-8 contributed to the failure of antibiotic treatment and consequently death of the patient.
authors.affiliation: Faculté des Sciences de Bizerte, Université de Carthage, Jarzouna 7021, Tunisia; Laboratoire de Microorganismes et Biomolécules Actives, Faculté des Sciences de Tunis, Université de Tunis El Manar, El Manar II 2098, Tunisia.
authors.forename: Rym
authors.initials: R
authors.lastname: Ouertani
authors.affiliation: CHU Reims, Hôpital Robert Debré, Laboratoire de Bactériologie-Virologie-Hygiène, Reims F-51092, France; UFR Médecine, SFR CAP-Santé, EA 4687, Université de Reims Champagne-Ardenne, Reims F-51092, France.
authors.forename: Anne
authors.initials: A
authors.lastname: Limelette
authors.affiliation: CHU Reims, Hôpital Robert Debré, Laboratoire de Bactériologie-Virologie-Hygiène, Reims F-51092, France; UFR Médecine, SFR CAP-Santé, EA 4687, Université de Reims Champagne-Ardenne, Reims F-51092, France.
authors.forename: Thomas
authors.initials: T
authors.lastname: Guillard
authors.affiliation: CHU Reims, Hôpital Robert Debré, Laboratoire de Bactériologie-Virologie-Hygiène, Reims F-51092, France.
authors.forename: Lucien
authors.initials: L
authors.lastname: Brasme
authors.affiliation: Centre Hospitalo-universitaire de Chirurgie Orthopédique et Traumatologique, Hôpital Régionale de Kasserine, Université de Sousse, Kasserine 1200, Tunisia.
authors.forename: Yahia
authors.initials: Y
authors.lastname: Jridi
authors.affiliation: Laboratoire de Microbiologie, Hôpital Militaire de Tunis, Montfleury, Tunis 1089, Tunisia.
authors.forename: Farouk
authors.initials: F
authors.lastname: Barguellil
authors.affiliation: Department of Environmental Health, Faculty of Public Health, Benghazi University, Benghazi, Libya; Al Abanos Medical Co. Al Seraj, Tripoli, Libya.
authors.forename: Allaaeddin
authors.initials: A
authors.lastname: El Salabi
authors.affiliation: CHU Reims, Hôpital Robert Debré, Laboratoire de Bactériologie-Virologie-Hygiène, Reims F-51092, France; UFR Médecine, SFR CAP-Santé, EA 4687, Université de Reims Champagne-Ardenne, Reims F-51092, France.
authors.forename: Christophe
authors.initials: C
authors.lastname: de Champs
authors.affiliation: Laboratoire de Microorganismes et Biomolécules Actives, Faculté des Sciences de Tunis, Université de Tunis El Manar, El Manar II 2098, Tunisia; Université de Carthage, Institut Supérieur des Sciences et Technologies de ľEnvironnement de Borj-Cedria, Technopôle de Borj-Cedria, BP-1003, Hammam-Lif 2050, Tunisia. Electronic address: chdoula77@yahoo.fr.
authors.forename: Chedly
authors.initials: C
authors.lastname: Chouchani
mesh_terms: D015373:Bacterial Typing Techniques; D003428:Cross Infection / Q000382:microbiology*; D024901:Drug Resistance, Multiple, Bacterial; D005798:Genes, Bacterial; D006801:Humans; D007710:Klebsiella Infections / Q000453:epidemiology*; D007711:Klebsiella pneumoniae / Q000201:enzymology / Q000235:genetics* / Q000302:isolation & purification; D058885:Multilocus Sequence Typing; D014416:Tunisia; D014774:Virulence; D001618:beta-Lactamases / Q000235:genetics*
chemical_list: D001618:beta-Lactamases
keywords: Carbapenemase; Inc plasmid; Klebsiella pneumoniae; OXA-48; VEB-8