Development and validation of a rapid real-time PCR based method for the specific detection of Salmonella on fresh meat

Sheila McGuinness; Evonne McCabe; Edel O'Regan; Anthony Dolan; Geraldine Duffy; Catherine Burgess; Seamus Fanning; Thomas Barry; Justin O'Grady

doi:10.1016/j.meatsci.2009.07.004

Development and validation of a rapid real-time PCR based method for the specific detection of Salmonella on fresh meat

Sheila McGuinness, Evonne McCabe, Edel O'Regan, Anthony Dolan, Geraldine Duffy, Catherine Burgess, Seamus Fanning, Thomas Barry, Justin O'Grady

Infectious Disease

Research output: Contribution to a Journal (Peer & Non Peer) › Article › peer-review

44 Citations (Scopus)

Abstract

In this study, a combined enrichment/real-time PCR method for the rapid detection of Salmonella on fresh meat carcasses, was designed, developed and validated in-house following requirements outlined in ISO 16140:2003. The method included an 18 h non-selective enrichment in buffered peptone water (BPW) and a 6 h selective enrichment in Rappaport Vasilliadis Soya (RVS) broth, based on the traditional culture method, ISO 6579:2002. The real-time PCR assay included an internal amplification control (IAC), was 100% specific and was sensitive to one cell equivalent. The alternative method was validated against the traditional culture method and relative accuracy of 94.9%, sensitivity of 94.7% and specificity of 100% were determined using 150 fresh meat carcass swabs. This alternative method had a detection limit of 1-10 CFU/100 cm² for fresh meat carcass swabs and was performed in 26 h. Following further inter-laboratory studies, this alternative method could be suitable for implementation in testing laboratories for the analysis of carcass swabs.

Original language	English
Pages (from-to)	555-562
Number of pages	8
Journal	Meat Science
Volume	83
Issue number	3
DOIs	https://doi.org/10.1016/j.meatsci.2009.07.004
Publication status	Published - Nov 2009

Keywords

Detection
Fresh meat
IAC
Real-time PCR
Salmonella
ssrA Gene

UN SDGs

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

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10.1016/j.meatsci.2009.07.004

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title = "Development and validation of a rapid real-time PCR based method for the specific detection of Salmonella on fresh meat",

abstract = "In this study, a combined enrichment/real-time PCR method for the rapid detection of Salmonella on fresh meat carcasses, was designed, developed and validated in-house following requirements outlined in ISO 16140:2003. The method included an 18 h non-selective enrichment in buffered peptone water (BPW) and a 6 h selective enrichment in Rappaport Vasilliadis Soya (RVS) broth, based on the traditional culture method, ISO 6579:2002. The real-time PCR assay included an internal amplification control (IAC), was 100\% specific and was sensitive to one cell equivalent. The alternative method was validated against the traditional culture method and relative accuracy of 94.9\%, sensitivity of 94.7\% and specificity of 100\% were determined using 150 fresh meat carcass swabs. This alternative method had a detection limit of 1-10 CFU/100 cm2 for fresh meat carcass swabs and was performed in 26 h. Following further inter-laboratory studies, this alternative method could be suitable for implementation in testing laboratories for the analysis of carcass swabs.",

keywords = "Detection, Fresh meat, IAC, Real-time PCR, Salmonella, ssrA Gene",

author = "Sheila McGuinness and Evonne McCabe and Edel O'Regan and Anthony Dolan and Geraldine Duffy and Catherine Burgess and Seamus Fanning and Thomas Barry and Justin O'Grady",

year = "2009",

month = nov,

doi = "10.1016/j.meatsci.2009.07.004",

language = "English",

volume = "83",

pages = "555--562",

journal = "Meat Science",

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T1 - Development and validation of a rapid real-time PCR based method for the specific detection of Salmonella on fresh meat

AU - McGuinness, Sheila

AU - McCabe, Evonne

AU - O'Regan, Edel

AU - Dolan, Anthony

AU - Duffy, Geraldine

AU - Burgess, Catherine

AU - Fanning, Seamus

AU - Barry, Thomas

AU - O'Grady, Justin

PY - 2009/11

Y1 - 2009/11

N2 - In this study, a combined enrichment/real-time PCR method for the rapid detection of Salmonella on fresh meat carcasses, was designed, developed and validated in-house following requirements outlined in ISO 16140:2003. The method included an 18 h non-selective enrichment in buffered peptone water (BPW) and a 6 h selective enrichment in Rappaport Vasilliadis Soya (RVS) broth, based on the traditional culture method, ISO 6579:2002. The real-time PCR assay included an internal amplification control (IAC), was 100% specific and was sensitive to one cell equivalent. The alternative method was validated against the traditional culture method and relative accuracy of 94.9%, sensitivity of 94.7% and specificity of 100% were determined using 150 fresh meat carcass swabs. This alternative method had a detection limit of 1-10 CFU/100 cm2 for fresh meat carcass swabs and was performed in 26 h. Following further inter-laboratory studies, this alternative method could be suitable for implementation in testing laboratories for the analysis of carcass swabs.

AB - In this study, a combined enrichment/real-time PCR method for the rapid detection of Salmonella on fresh meat carcasses, was designed, developed and validated in-house following requirements outlined in ISO 16140:2003. The method included an 18 h non-selective enrichment in buffered peptone water (BPW) and a 6 h selective enrichment in Rappaport Vasilliadis Soya (RVS) broth, based on the traditional culture method, ISO 6579:2002. The real-time PCR assay included an internal amplification control (IAC), was 100% specific and was sensitive to one cell equivalent. The alternative method was validated against the traditional culture method and relative accuracy of 94.9%, sensitivity of 94.7% and specificity of 100% were determined using 150 fresh meat carcass swabs. This alternative method had a detection limit of 1-10 CFU/100 cm2 for fresh meat carcass swabs and was performed in 26 h. Following further inter-laboratory studies, this alternative method could be suitable for implementation in testing laboratories for the analysis of carcass swabs.

KW - Detection

KW - Fresh meat

KW - IAC

KW - Real-time PCR

KW - Salmonella

KW - ssrA Gene

UR - https://www.scopus.com/pages/publications/69549121615

U2 - 10.1016/j.meatsci.2009.07.004

DO - 10.1016/j.meatsci.2009.07.004

M3 - Article

SN - 0309-1740

VL - 83

SP - 555

EP - 562

JO - Meat Science

JF - Meat Science

IS - 3

ER -

Development and validation of a rapid real-time PCR based method for the specific detection of Salmonella on fresh meat

Abstract

Keywords

UN SDGs

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