Comparison of phenotypic methods for the detection of carbapenem non-susceptible Enterobacteriaceae

Gut Pathogens

Giorgio Palù²  Sara N Richter²  Antonietta Cavallaro¹  Ilaria Frasson²  Andrea Bartolini¹ 

[1] Padua Hospital, Microbiology and Virology Unit, via Giustiniani 2, Padua 35121, Italy;Department of Molecular Medicine, University of Padua, via Gabelli 63, Padua 35121, Italy

关键词: Enterobacteriaceae;    Molecular analysis;    Genotypic assay;    Phenotypic detection;    Carbapenemases;   

Others  :  817518
DOI  :  10.1186/1757-4749-6-13

 received in 2014-03-24, accepted in 2014-05-06,  发布年份 2014

【 摘 要 】

Background

Multidrug resistance and, in particular, carbapenem resistance is spreading worldwide at an alarming rate, comprehending a variety of bacterial species and causing both nosocomial and community acquired outbursts. Early and efficient detection of infected patients or colonized carriers are mandatory steps in infection control and prevention of multidrug resistance diffusion. The latest EUCAST guidelines for detection of carbapenemase-producing Enterobacteriaceae have set low clinical breakpoints to ensure the maximum detection sensitivity of positive samples. Current workflows involve an initial screening step for species and resistance pattern detection, followed by phenotypic and/or genotypic confirmation. The aim of the present study was to assess the efficiency of six widely used and validated phenotypic assays for the detection of carbapenemases/AmpC in Enterobacteriaceae, to estimate the best workflow in the routine characterization of Enterobacteriaceae isolates.

Methods

A panel of 108 non-repetitive Enterobacteriaceae isolates with reduced susceptibility to carbapenems was analyzed by means of 1) Modified Hodge Test, 2) Metallo Beta Lactamase Etest, 3) Double disk test with EDTA, 4) Rosco Diagnostica KPC and MBL confirm kit (RDCK™), 5) AmpC Etest and 6) Cloxacillin inhibition test. Confirmation and validation of results was achieved by genotypic analysis.

Results

The most accurate identification of resistance determinants was obtained with the combined disc test (Rosco Diagnostica KPC and MBL confirm kit) which had to be coupled with the cloxacillin inhibition test for correct detection of AmpC enzymes. However, in general, phenotypic tests failed to characterize isolates harboring multiple carbapenem resistance determinants, which were successfully assessed only by PCR-based analysis.

Conclusions

To detect and control the spread of pathogens with complicated resistance patterns, both optimized phenotypic analysis (i.e. Rosco Diagnostica KPC and MBL confirm kit coupled with the cloxacillin inhibition test) and genotypic assays are recommended in the routine diagnostic of clinical laboratories.

【 授权许可】

   
2014 Bartolini et al.; licensee BioMed Central Ltd.

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