Detection and Occurrence of Plasmid-Mediated AmpC in Highly Resistant Gram-Negative Rods

E.A. Reuland, J.P. Hays, D.M.C. de Jongh, E. Abdelrehim, I. Willemsen, J.A.J.W. Kluytmans, P.H.M. Savelkoul, C.M.J.E. Vandenbroucke-Grauls, N. al Naiemi

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Abstract

Objectives: The aim of this study was to compare the current screening methods and to evaluate confirmation tests for phenotypic plasmidal AmpC (pAmpC) detection. Methods: For this evaluation we used 503 Enterobacteriaceae from 18 Dutch hospitals and 21 isolates previously confirmed to be pAmpC positive. All isolates were divided into three groups: isolates with 1) reduced susceptibility to ceftazidime and/or cefotaxime; 2) reduced susceptibility to cefoxitin; 3) reduced susceptibility to ceftazidime and/or cefotaxime combined with reduced susceptibility to cefoxitin. Two disk-based tests, with cloxacillin or boronic acid as inhibitor, and Etest with cefotetan-cefotetan/cloxacillin were used for phenotypic AmpC confirmation. Finally, presence of pAmpC genes was tested by multiplex and singleplex PCR. Results: We identified 13 pAmpC producing Enterobacteriaceae isolates among the 503 isolates (2.6%): 9 CMY-2, 3 DHA-1 and 1 ACC-1 type in E. coli isolates. The sensitivity and specificity of reduced susceptibility to ceftazidime and/or cefotaxime in combination with cefoxitin was 97% (33/34) and 90% (289/322) respectively. The disk-based test with cloxacillin showed the best performance as phenotypic confirmation method for AmpC production. Conclusions: For routine phenotypic detection of pAmpC the screening for reduced susceptibility to third generation cephalosporins combined with reduced susceptibility to cefoxitin is recommended. Confirmation via a combination disk diffusion test using cloxacillin is the best phenotypic option. The prevalence found is worrisome, since, due to their plasmidal location, pAmpC genes may spread further and increase in prevalence.
Original languageEnglish
Article numbere91396
JournalPLOS ONE
Volume9
Issue number3
DOIs
Publication statusPublished - 1 Jan 2014

Cite this

Reuland, E. A., Hays, J. P., de Jongh, D. M. C., Abdelrehim, E., Willemsen, I., Kluytmans, J. A. J. W., ... al Naiemi, N. (2014). Detection and Occurrence of Plasmid-Mediated AmpC in Highly Resistant Gram-Negative Rods. PLOS ONE, 9(3), [e91396]. https://doi.org/10.1371/journal.pone.0091396
Reuland, E.A. ; Hays, J.P. ; de Jongh, D.M.C. ; Abdelrehim, E. ; Willemsen, I. ; Kluytmans, J.A.J.W. ; Savelkoul, P.H.M. ; Vandenbroucke-Grauls, C.M.J.E. ; al Naiemi, N. / Detection and Occurrence of Plasmid-Mediated AmpC in Highly Resistant Gram-Negative Rods. In: PLOS ONE. 2014 ; Vol. 9, No. 3.
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title = "Detection and Occurrence of Plasmid-Mediated AmpC in Highly Resistant Gram-Negative Rods",
abstract = "Objectives: The aim of this study was to compare the current screening methods and to evaluate confirmation tests for phenotypic plasmidal AmpC (pAmpC) detection. Methods: For this evaluation we used 503 Enterobacteriaceae from 18 Dutch hospitals and 21 isolates previously confirmed to be pAmpC positive. All isolates were divided into three groups: isolates with 1) reduced susceptibility to ceftazidime and/or cefotaxime; 2) reduced susceptibility to cefoxitin; 3) reduced susceptibility to ceftazidime and/or cefotaxime combined with reduced susceptibility to cefoxitin. Two disk-based tests, with cloxacillin or boronic acid as inhibitor, and Etest with cefotetan-cefotetan/cloxacillin were used for phenotypic AmpC confirmation. Finally, presence of pAmpC genes was tested by multiplex and singleplex PCR. Results: We identified 13 pAmpC producing Enterobacteriaceae isolates among the 503 isolates (2.6{\%}): 9 CMY-2, 3 DHA-1 and 1 ACC-1 type in E. coli isolates. The sensitivity and specificity of reduced susceptibility to ceftazidime and/or cefotaxime in combination with cefoxitin was 97{\%} (33/34) and 90{\%} (289/322) respectively. The disk-based test with cloxacillin showed the best performance as phenotypic confirmation method for AmpC production. Conclusions: For routine phenotypic detection of pAmpC the screening for reduced susceptibility to third generation cephalosporins combined with reduced susceptibility to cefoxitin is recommended. Confirmation via a combination disk diffusion test using cloxacillin is the best phenotypic option. The prevalence found is worrisome, since, due to their plasmidal location, pAmpC genes may spread further and increase in prevalence.",
author = "E.A. Reuland and J.P. Hays and {de Jongh}, D.M.C. and E. Abdelrehim and I. Willemsen and J.A.J.W. Kluytmans and P.H.M. Savelkoul and C.M.J.E. Vandenbroucke-Grauls and {al Naiemi}, N.",
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Reuland, EA, Hays, JP, de Jongh, DMC, Abdelrehim, E, Willemsen, I, Kluytmans, JAJW, Savelkoul, PHM, Vandenbroucke-Grauls, CMJE & al Naiemi, N 2014, 'Detection and Occurrence of Plasmid-Mediated AmpC in Highly Resistant Gram-Negative Rods', PLOS ONE, vol. 9, no. 3, e91396. https://doi.org/10.1371/journal.pone.0091396

Detection and Occurrence of Plasmid-Mediated AmpC in Highly Resistant Gram-Negative Rods. / Reuland, E.A.; Hays, J.P.; de Jongh, D.M.C.; Abdelrehim, E.; Willemsen, I.; Kluytmans, J.A.J.W.; Savelkoul, P.H.M.; Vandenbroucke-Grauls, C.M.J.E.; al Naiemi, N.

In: PLOS ONE, Vol. 9, No. 3, e91396, 01.01.2014.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Detection and Occurrence of Plasmid-Mediated AmpC in Highly Resistant Gram-Negative Rods

AU - Reuland, E.A.

AU - Hays, J.P.

AU - de Jongh, D.M.C.

AU - Abdelrehim, E.

AU - Willemsen, I.

AU - Kluytmans, J.A.J.W.

AU - Savelkoul, P.H.M.

AU - Vandenbroucke-Grauls, C.M.J.E.

AU - al Naiemi, N.

PY - 2014/1/1

Y1 - 2014/1/1

N2 - Objectives: The aim of this study was to compare the current screening methods and to evaluate confirmation tests for phenotypic plasmidal AmpC (pAmpC) detection. Methods: For this evaluation we used 503 Enterobacteriaceae from 18 Dutch hospitals and 21 isolates previously confirmed to be pAmpC positive. All isolates were divided into three groups: isolates with 1) reduced susceptibility to ceftazidime and/or cefotaxime; 2) reduced susceptibility to cefoxitin; 3) reduced susceptibility to ceftazidime and/or cefotaxime combined with reduced susceptibility to cefoxitin. Two disk-based tests, with cloxacillin or boronic acid as inhibitor, and Etest with cefotetan-cefotetan/cloxacillin were used for phenotypic AmpC confirmation. Finally, presence of pAmpC genes was tested by multiplex and singleplex PCR. Results: We identified 13 pAmpC producing Enterobacteriaceae isolates among the 503 isolates (2.6%): 9 CMY-2, 3 DHA-1 and 1 ACC-1 type in E. coli isolates. The sensitivity and specificity of reduced susceptibility to ceftazidime and/or cefotaxime in combination with cefoxitin was 97% (33/34) and 90% (289/322) respectively. The disk-based test with cloxacillin showed the best performance as phenotypic confirmation method for AmpC production. Conclusions: For routine phenotypic detection of pAmpC the screening for reduced susceptibility to third generation cephalosporins combined with reduced susceptibility to cefoxitin is recommended. Confirmation via a combination disk diffusion test using cloxacillin is the best phenotypic option. The prevalence found is worrisome, since, due to their plasmidal location, pAmpC genes may spread further and increase in prevalence.

AB - Objectives: The aim of this study was to compare the current screening methods and to evaluate confirmation tests for phenotypic plasmidal AmpC (pAmpC) detection. Methods: For this evaluation we used 503 Enterobacteriaceae from 18 Dutch hospitals and 21 isolates previously confirmed to be pAmpC positive. All isolates were divided into three groups: isolates with 1) reduced susceptibility to ceftazidime and/or cefotaxime; 2) reduced susceptibility to cefoxitin; 3) reduced susceptibility to ceftazidime and/or cefotaxime combined with reduced susceptibility to cefoxitin. Two disk-based tests, with cloxacillin or boronic acid as inhibitor, and Etest with cefotetan-cefotetan/cloxacillin were used for phenotypic AmpC confirmation. Finally, presence of pAmpC genes was tested by multiplex and singleplex PCR. Results: We identified 13 pAmpC producing Enterobacteriaceae isolates among the 503 isolates (2.6%): 9 CMY-2, 3 DHA-1 and 1 ACC-1 type in E. coli isolates. The sensitivity and specificity of reduced susceptibility to ceftazidime and/or cefotaxime in combination with cefoxitin was 97% (33/34) and 90% (289/322) respectively. The disk-based test with cloxacillin showed the best performance as phenotypic confirmation method for AmpC production. Conclusions: For routine phenotypic detection of pAmpC the screening for reduced susceptibility to third generation cephalosporins combined with reduced susceptibility to cefoxitin is recommended. Confirmation via a combination disk diffusion test using cloxacillin is the best phenotypic option. The prevalence found is worrisome, since, due to their plasmidal location, pAmpC genes may spread further and increase in prevalence.

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VL - 9

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Reuland EA, Hays JP, de Jongh DMC, Abdelrehim E, Willemsen I, Kluytmans JAJW et al. Detection and Occurrence of Plasmid-Mediated AmpC in Highly Resistant Gram-Negative Rods. PLOS ONE. 2014 Jan 1;9(3). e91396. https://doi.org/10.1371/journal.pone.0091396