Combination of IMP-4 metallo-β-lactamase production and porin deficiency causes carbapenem resistance in a Klebsiella oxytoca clinical isolate

• Published in: Diagnostic microbiology and infectious disease 2009-10, Vol.65 (2), p.163-167
• Main Authors: Chen, Li-rong, Zhou, Hong-wei, Cai, Jia-chang, Zhang, Rong, Chen, Gong-xiang
• Format: Article
• Language: English
• Subjects: Anti-Bacterial Agents - pharmacology; Bacterial Proteins - analysis; Bacteriology; beta-Lactam Resistance; beta-Lactamases - chemistry; beta-Lactamases - genetics; beta-Lactamases - metabolism; Biological and medical sciences; Carbapenem resistance; Carbapenems - pharmacology; China; Class 1 integrons; Conjugation, Genetic; Electrophoresis, Polyacrylamide Gel; Escherichia coli - genetics; Fundamental and applied biological sciences. Psychology; Gene Transfer, Horizontal; Hospitals; Humans; Infectious Disease; Infectious diseases; Internal Medicine; Isoelectric Focusing; Klebsiella Infections - microbiology; Klebsiella oxytoca; Klebsiella oxytoca - drug effects; Klebsiella oxytoca - enzymology; Klebsiella oxytoca - genetics; Klebsiella oxytoca - isolation & purification; Medical sciences; Metallo-β-lactamases; Microbial Sensitivity Tests; Microbiology; Miscellaneous; Plasmids - analysis; porin; Porins - deficiency; Sequence Analysis, DNA
• ISSN: 0732-8893; 1879-0070
• DOI: 10.1016/j.diagmicrobio.2009.07.002

Abstract This study shows for the first time the mechanism of carbapenem resistance of a Klebsiella oxytoca clinical isolate ZC101 recovered from a Zhejiang University Hospital in Hangzhou, China. MIC values of imipenem, meropenem, and ertapenem for K. oxytoca ZC101 were 16, 16, and 128 μg/mL, respectively. Conjugation experiments demonstrated the transferability of a resistance determinant from K. oxytoca ZC101 to Escherichia coli EC600. Results from isoelectric focusing, polymerase chain reactions, and DNA sequencing confirmed that K. oxytoca ZC101 produced IMP-4 metallo-β-lactamase (MBL) and CTX-M-14 extended-spectrum β-lactamase, whereas E. coli transconjugant only produced the IMP-4. Amplification of integron revealed that blaIMP-4 gene is located within a class I integron that was carried in a plasmid approximately 55 kb in size. Sodium dodecyl sulfate polyacrylamide gel electrophoresis profiling of outer membrane proteins of K. oxytoca ZC101 indicated lack of expression of the OmpK36 porin. DNA sequence analysis of ompK 36 gene of K. oxytoca ZC101 showed the gene was disrupted by an insertion sequence IS5. In all, the results show that plasmid-mediated IMP-4 MBL production combined with the loss of OmpK36 porin caused the resistance in K. oxytoca ZC101 to carbapenems.