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Volume 46, Number 1, February 2013

Characterization of the modified Hodge test-positive isolates ofEnterobacteriaceae in Taiwan

 


Kuei-Hsiang Hung, Jing-Jou Yan, Jang-Jih Lu, Hung-Mo Chen, Jiunn-Jong Wu


Received: July 25, 2011    Revised: November 6, 2011    Accepted: December 19, 2011   

 

Corresponding author:

Jiunn-Jong Wu, Department of Medical Laboratory Science and Biotechnology, College of Medicine, National Cheng Kung University, Number 1 University Road, Tainan 70101, Taiwan.



 

Background and purpose: 

The modified Hodge test is a phenotypic test to detect KPC-type carbapenemase producers among Enterobacteriaceae, as recommended by the Clinical Laboratory Standards Institute. However, false positive results were reported. In this study, we aimed to large-scale investigate the characterization of the modified Hodge test-positive isolates of Enterobacteriaceaecollected between 2006 and 2010 in Taiwan.



 

Methods:

Fifty-six isolates, including 24 Enterobacter cloacae, 17 Escherichia coli, 10 Klebsiella pneumoniae, and 5 Citrobacter freundii, tested positive with the modified Hodge test. The in vitro activities of 10 antimicrobial agents were determined by the agar dilution method. Boronic acid combined-disk test was used to further confirm the KPC producers. Phenotype of ESBL, AmpC, class B carbapenemases, and profile of outer membrane proteins were investigated by the confirmatory test, boronic acid disk method, 2-mercaptopropionic acid double-disk method, and urea/sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), respectively. β-lactamase genes were examined by PCR and sequencing.



 

Results:

These isolates were resistant to ceftazidime (100%), aztreonam (82.1%), ertapenem (64.3%), gentamicin (53.6%), ciprofloxacin (50%), levofloxacin (48.2%), cefepime (19.6%), imipenem (16.1%), meropenem (12.5%), and amikacin (8.9%). Phenotypic testing among isolates revealed the production of ESBLs, metallo-β-lactamases (MBLs), and AmpC in 10 (17.9%), 16 (28.6%), and 12 (44.4%) isolates, respectively. Carbapenemase and non-carbapenemase β-lactamase genesblaTEM-1blaSHVblaCTX-MblaIMP-8blaCMY-2, and blaDHA-1 were found in 32 (57.1%), 19 (33.9%), 4 (7.1%), 16 (28.6%), 14 (25%), and 5 (8.9%) of the strains, respectively. No class A and D carbapenemase genes were detected. Outer membrane protein profile showed obviously decreased expression in 49 (87.5%) isolates with positive result of modified Hodge test.



 

Conclusion:

Our data show that ESBLs, AmpC, and imipenemase-8 (IMP-8) carbapenemase coupled with decreased expression of outer membrane protein were prevalent in Enterobacteriaceae isolates testing positive for the modified Hodge test in Taiwan.



 

Key words:

AmpCCarbapenemaseESBLModified Hodge testOuter membrane protein