In-House Validation of Multiplex PCR for Simultaneous Detection of Shiga Toxin-Producing Escherichia coli, Listeria monocytogenes and Salmonella spp. in Raw Meats

Chanokchon Jaroenporn; Wannakarn Supawasit; Damkerng Bundidamorn; Pathima Udompijitkul; Anunchai Assawamakin; Sudsai Trevanich

doi:10.3390/foods11111557

In-House Validation of Multiplex PCR for Simultaneous Detection of Shiga Toxin-Producing Escherichia coli, Listeria monocytogenes and Salmonella spp. in Raw Meats

Chanokchon Jaroenporn
, Wannakarn Supawasit
, Damkerng Bundidamorn
, Pathima Udompijitkul
, Anunchai Assawamakin
, Sudsai Trevanich

Department of Pharmacology

Kasetsart University

Research output: Contribution to journal › Article › peer-review

1 Citation (Scopus)

Abstract

The aim of the study was to perform in-house validation of the developed multiplex PCR (mPCR)-based alternative method to detect Shiga toxin-producing Escherichia coli (STEC), Listeria monocytogenes (L. monocytogenes) and Salmonella spp. in raw meats following the ISO 16140-2: 2016. A comparative study of the developed mPCR against the Bacteriological Analytical Manual (BAM) method was evaluated for inclusivity and exclusivity, sensitivity and the relative level of detection (RLOD). Inclusivity levels for each target bacterium were all 100%, while exclusivity for non-target bacteria was 100%. The sensitivity of the developed mPCR was calculated based on the analysis of 72 samples of raw meat. The sensitivity of the developed mPCR was 100%. The RLOD values of the developed mPCR for STEC, L. monocytogenes and Salmonella spp. were 0.756, 1.170 and 1.000, respectively. The developed mPCR showed potential as a tool for the fast, specific and sensitive detection of the three bacteria in the raw meat industry.

Original language	English
Article number	1557
Journal	Foods
Volume	11
Issue number	11
DOIs	https://doi.org/10.3390/foods11111557
Publication status	Published - 1 Jun 2022

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being
SDG 7 Affordable and Clean Energy
SDG 9 Industry, Innovation, and Infrastructure
SDG 12 Responsible Consumption and Production

Keywords

Listeria monocytogenes
Multiplex PCR
Salmonella spp
Shiga toxin-producing Escherichia coli
validation

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10.3390/foods11111557

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title = "In-House Validation of Multiplex PCR for Simultaneous Detection of Shiga Toxin-Producing Escherichia coli, Listeria monocytogenes and Salmonella spp. in Raw Meats",

abstract = "The aim of the study was to perform in-house validation of the developed multiplex PCR (mPCR)-based alternative method to detect Shiga toxin-producing Escherichia coli (STEC), Listeria monocytogenes (L. monocytogenes) and Salmonella spp. in raw meats following the ISO 16140-2: 2016. A comparative study of the developed mPCR against the Bacteriological Analytical Manual (BAM) method was evaluated for inclusivity and exclusivity, sensitivity and the relative level of detection (RLOD). Inclusivity levels for each target bacterium were all 100\%, while exclusivity for non-target bacteria was 100\%. The sensitivity of the developed mPCR was calculated based on the analysis of 72 samples of raw meat. The sensitivity of the developed mPCR was 100\%. The RLOD values of the developed mPCR for STEC, L. monocytogenes and Salmonella spp. were 0.756, 1.170 and 1.000, respectively. The developed mPCR showed potential as a tool for the fast, specific and sensitive detection of the three bacteria in the raw meat industry.",

keywords = "Listeria monocytogenes, Multiplex PCR, Salmonella spp, Shiga toxin-producing Escherichia coli, validation",

author = "Chanokchon Jaroenporn and Wannakarn Supawasit and Damkerng Bundidamorn and Pathima Udompijitkul and Anunchai Assawamakin and Sudsai Trevanich",

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doi = "10.3390/foods11111557",

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AU - Jaroenporn, Chanokchon

AU - Supawasit, Wannakarn

AU - Bundidamorn, Damkerng

AU - Udompijitkul, Pathima

AU - Assawamakin, Anunchai

AU - Trevanich, Sudsai

PY - 2022/6/1

Y1 - 2022/6/1

N2 - The aim of the study was to perform in-house validation of the developed multiplex PCR (mPCR)-based alternative method to detect Shiga toxin-producing Escherichia coli (STEC), Listeria monocytogenes (L. monocytogenes) and Salmonella spp. in raw meats following the ISO 16140-2: 2016. A comparative study of the developed mPCR against the Bacteriological Analytical Manual (BAM) method was evaluated for inclusivity and exclusivity, sensitivity and the relative level of detection (RLOD). Inclusivity levels for each target bacterium were all 100%, while exclusivity for non-target bacteria was 100%. The sensitivity of the developed mPCR was calculated based on the analysis of 72 samples of raw meat. The sensitivity of the developed mPCR was 100%. The RLOD values of the developed mPCR for STEC, L. monocytogenes and Salmonella spp. were 0.756, 1.170 and 1.000, respectively. The developed mPCR showed potential as a tool for the fast, specific and sensitive detection of the three bacteria in the raw meat industry.

AB - The aim of the study was to perform in-house validation of the developed multiplex PCR (mPCR)-based alternative method to detect Shiga toxin-producing Escherichia coli (STEC), Listeria monocytogenes (L. monocytogenes) and Salmonella spp. in raw meats following the ISO 16140-2: 2016. A comparative study of the developed mPCR against the Bacteriological Analytical Manual (BAM) method was evaluated for inclusivity and exclusivity, sensitivity and the relative level of detection (RLOD). Inclusivity levels for each target bacterium were all 100%, while exclusivity for non-target bacteria was 100%. The sensitivity of the developed mPCR was calculated based on the analysis of 72 samples of raw meat. The sensitivity of the developed mPCR was 100%. The RLOD values of the developed mPCR for STEC, L. monocytogenes and Salmonella spp. were 0.756, 1.170 and 1.000, respectively. The developed mPCR showed potential as a tool for the fast, specific and sensitive detection of the three bacteria in the raw meat industry.

KW - Listeria monocytogenes

KW - Multiplex PCR

KW - Salmonella spp

KW - Shiga toxin-producing Escherichia coli

KW - validation

UR - https://www.scopus.com/pages/publications/85131552578

U2 - 10.3390/foods11111557

DO - 10.3390/foods11111557

M3 - Article

AN - SCOPUS:85131552578

SN - 2304-8158

VL - 11

JO - Foods

JF - Foods

IS - 11

M1 - 1557

ER -

In-House Validation of Multiplex PCR for Simultaneous Detection of Shiga Toxin-Producing Escherichia coli, Listeria monocytogenes and Salmonella spp. in Raw Meats

Abstract

UN SDGs

Keywords

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