MALDI-TOF MS based carbapenemase detection from culture isolates and from positive blood culture vials

BACKGROUND: Antibiotic resistance in bacteria leads to massive health problems. Incidence of carbapenem and multidrug resistance in Gram-negative bacteria are increasing globally and turn out to be a very urgent challenge in health care. Resistant bacteria play an important clinical role during hosp...

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Autores principales: Ghebremedhin, B., Halstenbach, A., Smiljanic, M., Kaase, M., Ahmad-Nejad, P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4736273/
https://www.ncbi.nlm.nih.gov/pubmed/26839024
http://dx.doi.org/10.1186/s12941-016-0120-x
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author Ghebremedhin, B.
Halstenbach, A.
Smiljanic, M.
Kaase, M.
Ahmad-Nejad, P.
author_facet Ghebremedhin, B.
Halstenbach, A.
Smiljanic, M.
Kaase, M.
Ahmad-Nejad, P.
author_sort Ghebremedhin, B.
collection PubMed
description BACKGROUND: Antibiotic resistance in bacteria leads to massive health problems. Incidence of carbapenem and multidrug resistance in Gram-negative bacteria are increasing globally and turn out to be a very urgent challenge in health care. Resistant bacteria play an important clinical role during hospital outbreaks as well as in sepsis. Rapid diagnostic tests are necessary to provide immediate information for antimicrobial treatment and infection control measures. METHODS: Our mass spectrometry-based assay was validated with 63 carbapenemase-producing Gram-negative bacterial isolates, and 35 carbapenem-resistant Gram-negative species with no carbapenemase production. These were analyzed from solid culture media and positive blood culture vials. After 4 h of incubation the carbapenemase products were analyzed with the MALDI-TOF MS. All the isolates were genotyped for carbapenemase genes by PCR and sequencing. RESULTS: For culture isolates the concordance of hydrolysis assay to genetic results was 98 % for OXA variants, KPC, VIM, IMP, GIM, and NDM. In contrast, only 14 of 29 Acinetobacter baumannii isolates carrying the OXA and NDM genes could be identified from blood culture. However, from blood culture vials our method allowed the detection of carbapenemases in 98 % of Pseudomonas and Enterobacteriaceae isolates harboring different genes. CONCLUSIONS: This MALDI-TOF MS–based assay permitted the detection of carbapenemases either from solid culture media (98–100 %) or blood culture vials (96 %) for all non-A. baumannii isolates within 4 h. In case of A. baumannii isolates the assay was highly sensitive for the detection of carbapenemases directly from solid culture media.
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spelling pubmed-47362732016-02-03 MALDI-TOF MS based carbapenemase detection from culture isolates and from positive blood culture vials Ghebremedhin, B. Halstenbach, A. Smiljanic, M. Kaase, M. Ahmad-Nejad, P. Ann Clin Microbiol Antimicrob Research BACKGROUND: Antibiotic resistance in bacteria leads to massive health problems. Incidence of carbapenem and multidrug resistance in Gram-negative bacteria are increasing globally and turn out to be a very urgent challenge in health care. Resistant bacteria play an important clinical role during hospital outbreaks as well as in sepsis. Rapid diagnostic tests are necessary to provide immediate information for antimicrobial treatment and infection control measures. METHODS: Our mass spectrometry-based assay was validated with 63 carbapenemase-producing Gram-negative bacterial isolates, and 35 carbapenem-resistant Gram-negative species with no carbapenemase production. These were analyzed from solid culture media and positive blood culture vials. After 4 h of incubation the carbapenemase products were analyzed with the MALDI-TOF MS. All the isolates were genotyped for carbapenemase genes by PCR and sequencing. RESULTS: For culture isolates the concordance of hydrolysis assay to genetic results was 98 % for OXA variants, KPC, VIM, IMP, GIM, and NDM. In contrast, only 14 of 29 Acinetobacter baumannii isolates carrying the OXA and NDM genes could be identified from blood culture. However, from blood culture vials our method allowed the detection of carbapenemases in 98 % of Pseudomonas and Enterobacteriaceae isolates harboring different genes. CONCLUSIONS: This MALDI-TOF MS–based assay permitted the detection of carbapenemases either from solid culture media (98–100 %) or blood culture vials (96 %) for all non-A. baumannii isolates within 4 h. In case of A. baumannii isolates the assay was highly sensitive for the detection of carbapenemases directly from solid culture media. BioMed Central 2016-02-02 /pmc/articles/PMC4736273/ /pubmed/26839024 http://dx.doi.org/10.1186/s12941-016-0120-x Text en © Ghebremedhin et al. 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Ghebremedhin, B.
Halstenbach, A.
Smiljanic, M.
Kaase, M.
Ahmad-Nejad, P.
MALDI-TOF MS based carbapenemase detection from culture isolates and from positive blood culture vials
title MALDI-TOF MS based carbapenemase detection from culture isolates and from positive blood culture vials
title_full MALDI-TOF MS based carbapenemase detection from culture isolates and from positive blood culture vials
title_fullStr MALDI-TOF MS based carbapenemase detection from culture isolates and from positive blood culture vials
title_full_unstemmed MALDI-TOF MS based carbapenemase detection from culture isolates and from positive blood culture vials
title_short MALDI-TOF MS based carbapenemase detection from culture isolates and from positive blood culture vials
title_sort maldi-tof ms based carbapenemase detection from culture isolates and from positive blood culture vials
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4736273/
https://www.ncbi.nlm.nih.gov/pubmed/26839024
http://dx.doi.org/10.1186/s12941-016-0120-x
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