Comparative Evaluation of the VITEK 2, Disk Diffusion, Etest, Broth Microdilution, and Agar Dilution Susceptibility Testing Methods for Colistin in Clinical Isolates, Including Heteroresistant Enterobacter cloacae and Acinetobacter baumannii Strains

Lo-Ten-Foe, Jerome R.; de Smet, Anne Marie G. A.; Diederen, Bram M. W.; Kluytmans, Jan A. J. W.; van Keulen, Peter H. J.

doi:10.1128/aac.01406-06

Cited by 242Antibiotic Resistance in BacteriaBacterial Identification and Susceptibility TestingAntibiotics Pharmacokinetics and Efficacy

Comparative Evaluation of the VITEK 2, Disk Diffusion, Etest, Broth Microdilution, and Agar Dilution Susceptibility Testing Methods for Colistin in Clinical Isolates, Including Heteroresistant Enterobacter cloacae and Acinetobacter baumannii Strains

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Lo-Ten-Foe, Jerome R., de Smet, Anne Marie G. A., Diederen, Bram M. W., Kluytmans, Jan A. J. W., van Keulen, Peter H. J.·2007·Antimicrobial Agents and Chemotherapy

DOI 10.1128/aac.01406-06PMC PMC2043258

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Abstract

Increasing antibiotic resistance in gram-negative bacteria has recently renewed interest in colistin as a therapeutic option. The increasing use of colistin necessitates the availability of rapid and reliable methods for colistin susceptibility testing. We compared seven methods of colistin susceptibility testing (disk diffusion, agar dilution on Mueller-Hinton [MH] and Isosensitest agar, Etest on MH and Isosensitest agar, broth microdilution, and VITEK 2) on 102 clinical isolates collected from patient materials during a selective digestive decontamination or selective oral decontamination trial in an intensive-care unit. Disk diffusion is an unreliable method to measure susceptibility to colistin. High error rates and low levels of reproducibility were observed in the disk diffusion test. The colistin Etest, agar dilution, and the VITEK 2 showed a high level of agreement with the broth microdilution reference method. Heteroresistance for colistin was observed in six Enterobacter cloacae isolates and in one Acinetobacter baumannii isolate. This is the first report of heteroresistance to colistin in E. cloacae isolates. Resistance to colistin in these isolates seemed to be induced upon exposure to colistin rather than being caused by stable mutations. Heteroresistant isolates could be detected in the broth microdilution, agar dilution, Etest, or disk diffusion test. The VITEK 2 displayed low sensitivity in the detection of heteroresistant subpopulations of E. cloacae . The VITEK 2 colistin susceptibility test can therefore be considered to be a reliable tool to determine susceptibility to colistin in isolates of genera that are known not to exhibit resistant subpopulations. In isolates of genera known to (occasionally) exhibit heteroresistance, an alternative susceptibility testing method capable of detecting heteroresistance should be used.

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E-test results significantly correlate with MIC values obtained through broth or agar dilution methods.
“studies have shown a significant correlation between E-test results and MIC values obtained through broth or agar dilution methods [69, 70].”
From Methods for screening and evaluation of antimicrobial activity: A review of protocols, advantages, and limitations (2024) · cites this paper as [69]
The Results (page not supplied) report relatively high agreement between Etest and broth microdilution for colistin susceptibility testing, with exceptions among heteroresistant isolates. They do not report a direct Etest–agar dilution comparison or a statistical test establishing that the correlation was significant. (LLM, conf=0.78)
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Authors: Lo-Ten-Foe, J. R., de Smet, A. M. G. A., Diederen, B. M. W., Kluytmans, J. A. J. W., & van Keulen, P. H. J.
Affiliations: Laboratory for Microbiology and Infection Control, Amphia Hospital, Breda, The Netherlands
Perioperative and Emergency Care, Utrecht Medical Center, Utrecht, The Netherlands
Amphia Ziekenhuis (NL)
ORCID: Kluytmans, Jan A. J. W.
Venue: Antimicrobial Agents and Chemotherapy
Publisher: American Society for Microbiology
Published: 2007-10
Volume: 51
Issue: 10
Pages: 3726-3730
ISSN: 0066-4804 (ISSN-L)
1098-6596
DOI: 10.1128/aac.01406-06
PMC: PMC2043258
OpenAlex: W2142419984
Cited by: 242
References: 22
Citations by year: 2026: 15 · 2025: 10 · 2024: 16 · 2023: 19 · 2022: 14 · 2021: 9 · 2020: 22 · 2019: 16 · 2018: 19 · 2017: 24 · 2016: 13 · 2015: 11 · 2014: 9 · 2013: 11 · 2012: 7
FWCI: 3.30
Type: journal article
Language: en
Open access: green
www.ncbi.nlm.nih.gov
Licence: journals.asm.org/non-commercial-tdm-license (tdm)
Locations: Antimicrobial Agents and Chemotherapy (publishedVersion)
PubMed Central (publishedVersion)
PubMed (publishedVersion)
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Medicine › Antibiotic Resistance in Bacteria
Life Sciences › Biochemistry, Genetics and Molecular Biology › Clinical Biochemistry › Bacterial Identification and Susceptibility Testing
Health Sciences › Medicine › Pharmacology › Antibiotics Pharmacokinetics and Efficacy
Keywords: colistin, colistin susceptibility testing, heteroresistance, Enterobacter cloacae, Acinetobacter baumannii, disk diffusion, E-test, broth microdilution, agar dilution, VITEK 2, selective decontamination of the digestive tract, intensive care unit, Gram-negative bacteria, antibiotic resistance
MeSH terms: Acinetobacter Infections, Agar, Anti-Bacterial Agents, Bacterial Infections, Colistin, Cross Infection, Culture Media, Enterobacteriaceae Infections, Humans, Intensive Care Units, Microbial Sensitivity Tests, Polymyxin B, Enterobacter cloacae, Drug Resistance, Multiple, Bacterial, Acinetobacter baumannii

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Comparative Evaluation of the VITEK 2, Disk Diffusion, Etest, Broth Microdilution, and Agar Dilution Susceptibility Testing Methods for Colistin in Clinical Isolates, Including Heteroresistant Enterobacter cloacae and Acinetobacter baumannii Strains

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