Impact of various killing methods on EMA/PMA-qPCR efficacy

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F60461373%3A22330%2F18%3A43914945" target="_blank" >RIV/60461373:22330/18:43914945 - isvavai.cz</a>

Alternative codes found

RIV/60461373:22340/18:43914945 RIV/60461373:22810/18:43914945

Result on the web

<a href="https://www.sciencedirect.com/science/article/pii/S0956713517304486" target="_blank" >https://www.sciencedirect.com/science/article/pii/S0956713517304486</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1016/j.foodcont.2017.09.013" target="_blank" >10.1016/j.foodcont.2017.09.013</a>

Result language

angličtina

Original language name

Impact of various killing methods on EMA/PMA-qPCR efficacy

Original language description

Although qPCR is indisputably very powerful tool for identification and quantification of various microorganisms, one of its main disadvantages is the inability to distinguish between signals originating from different sources (live, dead or compromised cells). Sample pre-treatment with DNA intercalating dyes such as ethidium monoazide (EMA) or propidium monoazide (PMA) is a widely used solution which helps to reduce this issue. Efficacy of this pre-treatment, however, depends on membrane permeability. Therefore in this work, a killing assay was performed, evaluating the impact of several cell inactivation techniques with various modes of action on EMA/PMA-qPCR results. Three main thermotolerant Campylobacter spp. (C. jejuni, C. coli and C. lari) in mixed suspension were inactivated by a panel of eight lethal and one sub-lethal procedures, which bacteria can encounter during their life span (e.g. food processing and storage, surface disinfection or laboratory maintenance in terms of sterilization or aseptic work), and afterwards were quantified using previously designed multiplex EMA/PMA-qPCR. It was shown that not only various inactivation mechanisms affect the efficacy of qPCR, but species-specific differences were apparent as well. C. lari was the most resistant species to EMA/PMA treatment, while C. coli seemed to be the most sensitive. Results of this killing assay might be generally helpful in every research where EMA/PMA-qPCR is used, as they clearly show that this pre-treatment technique cannot be blindly used for every sample without at least considering the way of the bacterial inactivation.

Czech name

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Czech description

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Type

J_imp - Article in a specialist periodical, which is included in the Web of Science database

CEP classification

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OECD FORD branch

10606 - Microbiology

Project

<a href="/en/project/GA17-15936S" target="_blank" >GA17-15936S: Interaction of nanoparticles modified by natural compounds with biofilms of pathogenic microorganisms</a><br>

Continuities

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

Publication year

2018

Confidentiality

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

Name of the periodical

Food control

ISSN

0956-7135

e-ISSN

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Volume of the periodical

85

Issue of the periodical within the volume

MAR

Country of publishing house

BE - BELGIUM

Number of pages

6

Pages from-to

23-28

UT code for WoS article

000419409000004

EID of the result in the Scopus database

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