Typing of Enterobacteriaceae by ERIC-PCR, REP-PCR and MALDI-TOF MS methods

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F60461373%3A22330%2F15%3A43901505" target="_blank" >RIV/60461373:22330/15:43901505 - isvavai.cz</a>

Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

Typing of Enterobacteriaceae by ERIC-PCR, REP-PCR and MALDI-TOF MS methods

Popis výsledku v původním jazyce

The fast and reliable fingerprinting methods for typing of Enterobacteriaceae are widely required in food microbiology for tracking sources and ways of food contamination. ERIC-PCR and REP-PCR genotyping use as markers specific DNA sequences (enterobacterial repetitive intergenic consensus sequences or repetitive extragenic palindronic sequences, respectively) and the changes of their dispersal, detactable by methods. In comparison MALDI-TOF MS fenotyping uses the conformity of protein profiles. As used typing markers mirror the differences among strains only partially, also compared to other features as the antibiotics resistance and biofilm formation, their combination could propose more precise approach. The aim was to compare the discriminatory power of ERIC-PCR, REP-PCR and MALDI-TOF MS fingerprinting methods for different Enterobacteriaceae isolates, also with respect to their antibiotics resistance and biofilm formation. Fifty six isolates of Enterobacteriaceae, including genera Escherichia, Enterobacter, Klebsiella, Serratia and Raoultella, isolated from food and clinical sources in Czech Republic in 2007-2014, were genotyped by the same ERIC-PCR and REP-PCR methods (maximal length of analysed fragments 3000 bp). MALDI-TOF MS typing was done by the ethanol-formic acid extraction in Bruker Autoflex Speed MALDI-TOF mass spectrometer. The resistance for 12 antibiotics and the ability to form biofilm in different media at 25 oC were tested. The groups of genus and species specific markers in all methods were found, but the discriminatory power of methods within species was different. The applicability of MALDI-TOF MS typing for further discrimination within isolates of congruent ERIC-PCR and REP-PCR profiles depends on the used method of cluster analysis.

Název v anglickém jazyce

Typing of Enterobacteriaceae by ERIC-PCR, REP-PCR and MALDI-TOF MS methods

Popis výsledku anglicky

The fast and reliable fingerprinting methods for typing of Enterobacteriaceae are widely required in food microbiology for tracking sources and ways of food contamination. ERIC-PCR and REP-PCR genotyping use as markers specific DNA sequences (enterobacterial repetitive intergenic consensus sequences or repetitive extragenic palindronic sequences, respectively) and the changes of their dispersal, detactable by methods. In comparison MALDI-TOF MS fenotyping uses the conformity of protein profiles. As used typing markers mirror the differences among strains only partially, also compared to other features as the antibiotics resistance and biofilm formation, their combination could propose more precise approach. The aim was to compare the discriminatory power of ERIC-PCR, REP-PCR and MALDI-TOF MS fingerprinting methods for different Enterobacteriaceae isolates, also with respect to their antibiotics resistance and biofilm formation. Fifty six isolates of Enterobacteriaceae, including genera Escherichia, Enterobacter, Klebsiella, Serratia and Raoultella, isolated from food and clinical sources in Czech Republic in 2007-2014, were genotyped by the same ERIC-PCR and REP-PCR methods (maximal length of analysed fragments 3000 bp). MALDI-TOF MS typing was done by the ethanol-formic acid extraction in Bruker Autoflex Speed MALDI-TOF mass spectrometer. The resistance for 12 antibiotics and the ability to form biofilm in different media at 25 oC were tested. The groups of genus and species specific markers in all methods were found, but the discriminatory power of methods within species was different. The applicability of MALDI-TOF MS typing for further discrimination within isolates of congruent ERIC-PCR and REP-PCR profiles depends on the used method of cluster analysis.

Druh

O - Ostatní výsledky

CEP obor

EE - Mikrobiologie, virologie

OECD FORD obor

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Projekt

<a href="/cs/project/QJ1210300" target="_blank" >QJ1210300: Systémy jištění kvality a bezpečnosti mlékárenských výrobků vhodnými metodami aplikovatelnými v praxi</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2015

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů