A qPCR Assay to Detect and Quantify Shiga Toxin-Producing E. coli (STEC) in Cattle and on Farms: A Potential Predictive Tool for STEC Culture-Positive Farms
Shiga toxin-producing E. coli (STEC), of various serogroups harboring the intimin gene, form a serious threat to human health. They are asymptomatically carried by cattle. In this study, a quantitative real-time PCR (qPCR) method was developed as a molecular method to detect and quantify Shiga toxin...
Main Authors: | , , , , , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
MDPI AG
2014-03-01
|
Series: | Toxins |
Subjects: | |
Online Access: | http://www.mdpi.com/2072-6651/6/4/1201 |
id |
doaj-1c29d1678313439ba1ced749d53094bb |
---|---|
record_format |
Article |
spelling |
doaj-1c29d1678313439ba1ced749d53094bb2020-11-24T22:02:17ZengMDPI AGToxins2072-66512014-03-01641201122110.3390/toxins6041201toxins6041201A qPCR Assay to Detect and Quantify Shiga Toxin-Producing E. coli (STEC) in Cattle and on Farms: A Potential Predictive Tool for STEC Culture-Positive FarmsKaren Verstraete0Els Van Coillie1Hadewig Werbrouck2Stephanie Van Weyenberg3Lieve Herman4Jurgen Del-Favero5Peter De Rijk6Lieven De Zutter7Maria-Adelheid Joris8Marc Heyndrickx9Koen DeReu10Technology and Food Science Unit, Institute for Agricultural and Fisheries Research (ILVO), Brusselsesteenweg 370, Melle 9090, BelgiumTechnology and Food Science Unit, Institute for Agricultural and Fisheries Research (ILVO), Brusselsesteenweg 370, Melle 9090, BelgiumTechnology and Food Science Unit, Institute for Agricultural and Fisheries Research (ILVO), Brusselsesteenweg 370, Melle 9090, BelgiumTechnology and Food Science Unit, Institute for Agricultural and Fisheries Research (ILVO), Brusselsesteenweg 370, Melle 9090, BelgiumTechnology and Food Science Unit, Institute for Agricultural and Fisheries Research (ILVO), Brusselsesteenweg 370, Melle 9090, BelgiumApplied Molecular Genomics Group, Department of Molecular Genetics, Flemish Institute for Biotechnology (VIB), Universiteitsplein 1, Antwerpen 2610, BelgiumApplied Molecular Genomics Group, Department of Molecular Genetics, Flemish Institute for Biotechnology (VIB), Universiteitsplein 1, Antwerpen 2610, BelgiumDepartment of Veterinary Public Health and Food Safety, Ghent University, Salisburylaan 133, Merelbeke 9820, BelgiumDepartment of Veterinary Public Health and Food Safety, Ghent University, Salisburylaan 133, Merelbeke 9820, BelgiumTechnology and Food Science Unit, Institute for Agricultural and Fisheries Research (ILVO), Brusselsesteenweg 370, Melle 9090, BelgiumTechnology and Food Science Unit, Institute for Agricultural and Fisheries Research (ILVO), Brusselsesteenweg 370, Melle 9090, BelgiumShiga toxin-producing E. coli (STEC), of various serogroups harboring the intimin gene, form a serious threat to human health. They are asymptomatically carried by cattle. In this study, a quantitative real-time PCR (qPCR) method was developed as a molecular method to detect and quantify Shiga toxin genes stx1 and stx2 and the intimin gene eae. Subsequently, 59 fecal samples from six farms were tested using qPCR and a culture method as a reference. Three farms had contaminated animals as demonstrated by the culture method. Culture-positive farms showed moderate significantly higher stx prevalences than culture-negative farms (p = 0.05). This is the first study which showed preliminary results that qPCR can predict STEC farm contamination, with a specificity of 77% and a sensitivity of 83%, as compared with the culture method. Furthermore, the presence or quantity of stx genes in feces was not correlated to the isolation of STEC from the individual animal. Quantitative data thus did not add value to the results. Finally, the detection of both stx and eae genes within the same fecal sample or farm using qPCR was not correlated with the isolation of an eae-harboring STEC strain from the respective sample or farm using the culture method.http://www.mdpi.com/2072-6651/6/4/1201Shiga toxinE. colireal-time PCRcattlequantificationintiminscreeningfarmfecesisolation |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Karen Verstraete Els Van Coillie Hadewig Werbrouck Stephanie Van Weyenberg Lieve Herman Jurgen Del-Favero Peter De Rijk Lieven De Zutter Maria-Adelheid Joris Marc Heyndrickx Koen DeReu |
spellingShingle |
Karen Verstraete Els Van Coillie Hadewig Werbrouck Stephanie Van Weyenberg Lieve Herman Jurgen Del-Favero Peter De Rijk Lieven De Zutter Maria-Adelheid Joris Marc Heyndrickx Koen DeReu A qPCR Assay to Detect and Quantify Shiga Toxin-Producing E. coli (STEC) in Cattle and on Farms: A Potential Predictive Tool for STEC Culture-Positive Farms Toxins Shiga toxin E. coli real-time PCR cattle quantification intimin screening farm feces isolation |
author_facet |
Karen Verstraete Els Van Coillie Hadewig Werbrouck Stephanie Van Weyenberg Lieve Herman Jurgen Del-Favero Peter De Rijk Lieven De Zutter Maria-Adelheid Joris Marc Heyndrickx Koen DeReu |
author_sort |
Karen Verstraete |
title |
A qPCR Assay to Detect and Quantify Shiga Toxin-Producing E. coli (STEC) in Cattle and on Farms: A Potential Predictive Tool for STEC Culture-Positive Farms |
title_short |
A qPCR Assay to Detect and Quantify Shiga Toxin-Producing E. coli (STEC) in Cattle and on Farms: A Potential Predictive Tool for STEC Culture-Positive Farms |
title_full |
A qPCR Assay to Detect and Quantify Shiga Toxin-Producing E. coli (STEC) in Cattle and on Farms: A Potential Predictive Tool for STEC Culture-Positive Farms |
title_fullStr |
A qPCR Assay to Detect and Quantify Shiga Toxin-Producing E. coli (STEC) in Cattle and on Farms: A Potential Predictive Tool for STEC Culture-Positive Farms |
title_full_unstemmed |
A qPCR Assay to Detect and Quantify Shiga Toxin-Producing E. coli (STEC) in Cattle and on Farms: A Potential Predictive Tool for STEC Culture-Positive Farms |
title_sort |
qpcr assay to detect and quantify shiga toxin-producing e. coli (stec) in cattle and on farms: a potential predictive tool for stec culture-positive farms |
publisher |
MDPI AG |
series |
Toxins |
issn |
2072-6651 |
publishDate |
2014-03-01 |
description |
Shiga toxin-producing E. coli (STEC), of various serogroups harboring the intimin gene, form a serious threat to human health. They are asymptomatically carried by cattle. In this study, a quantitative real-time PCR (qPCR) method was developed as a molecular method to detect and quantify Shiga toxin genes stx1 and stx2 and the intimin gene eae. Subsequently, 59 fecal samples from six farms were tested using qPCR and a culture method as a reference. Three farms had contaminated animals as demonstrated by the culture method. Culture-positive farms showed moderate significantly higher stx prevalences than culture-negative farms (p = 0.05). This is the first study which showed preliminary results that qPCR can predict STEC farm contamination, with a specificity of 77% and a sensitivity of 83%, as compared with the culture method. Furthermore, the presence or quantity of stx genes in feces was not correlated to the isolation of STEC from the individual animal. Quantitative data thus did not add value to the results. Finally, the detection of both stx and eae genes within the same fecal sample or farm using qPCR was not correlated with the isolation of an eae-harboring STEC strain from the respective sample or farm using the culture method. |
topic |
Shiga toxin E. coli real-time PCR cattle quantification intimin screening farm feces isolation |
url |
http://www.mdpi.com/2072-6651/6/4/1201 |
work_keys_str_mv |
AT karenverstraete aqpcrassaytodetectandquantifyshigatoxinproducingecolistecincattleandonfarmsapotentialpredictivetoolforstecculturepositivefarms AT elsvancoillie aqpcrassaytodetectandquantifyshigatoxinproducingecolistecincattleandonfarmsapotentialpredictivetoolforstecculturepositivefarms AT hadewigwerbrouck aqpcrassaytodetectandquantifyshigatoxinproducingecolistecincattleandonfarmsapotentialpredictivetoolforstecculturepositivefarms AT stephanievanweyenberg aqpcrassaytodetectandquantifyshigatoxinproducingecolistecincattleandonfarmsapotentialpredictivetoolforstecculturepositivefarms AT lieveherman aqpcrassaytodetectandquantifyshigatoxinproducingecolistecincattleandonfarmsapotentialpredictivetoolforstecculturepositivefarms AT jurgendelfavero aqpcrassaytodetectandquantifyshigatoxinproducingecolistecincattleandonfarmsapotentialpredictivetoolforstecculturepositivefarms AT peterderijk aqpcrassaytodetectandquantifyshigatoxinproducingecolistecincattleandonfarmsapotentialpredictivetoolforstecculturepositivefarms AT lievendezutter aqpcrassaytodetectandquantifyshigatoxinproducingecolistecincattleandonfarmsapotentialpredictivetoolforstecculturepositivefarms AT mariaadelheidjoris aqpcrassaytodetectandquantifyshigatoxinproducingecolistecincattleandonfarmsapotentialpredictivetoolforstecculturepositivefarms AT marcheyndrickx aqpcrassaytodetectandquantifyshigatoxinproducingecolistecincattleandonfarmsapotentialpredictivetoolforstecculturepositivefarms AT koendereu aqpcrassaytodetectandquantifyshigatoxinproducingecolistecincattleandonfarmsapotentialpredictivetoolforstecculturepositivefarms AT karenverstraete qpcrassaytodetectandquantifyshigatoxinproducingecolistecincattleandonfarmsapotentialpredictivetoolforstecculturepositivefarms AT elsvancoillie qpcrassaytodetectandquantifyshigatoxinproducingecolistecincattleandonfarmsapotentialpredictivetoolforstecculturepositivefarms AT hadewigwerbrouck qpcrassaytodetectandquantifyshigatoxinproducingecolistecincattleandonfarmsapotentialpredictivetoolforstecculturepositivefarms AT stephanievanweyenberg qpcrassaytodetectandquantifyshigatoxinproducingecolistecincattleandonfarmsapotentialpredictivetoolforstecculturepositivefarms AT lieveherman qpcrassaytodetectandquantifyshigatoxinproducingecolistecincattleandonfarmsapotentialpredictivetoolforstecculturepositivefarms AT jurgendelfavero qpcrassaytodetectandquantifyshigatoxinproducingecolistecincattleandonfarmsapotentialpredictivetoolforstecculturepositivefarms AT peterderijk qpcrassaytodetectandquantifyshigatoxinproducingecolistecincattleandonfarmsapotentialpredictivetoolforstecculturepositivefarms AT lievendezutter qpcrassaytodetectandquantifyshigatoxinproducingecolistecincattleandonfarmsapotentialpredictivetoolforstecculturepositivefarms AT mariaadelheidjoris qpcrassaytodetectandquantifyshigatoxinproducingecolistecincattleandonfarmsapotentialpredictivetoolforstecculturepositivefarms AT marcheyndrickx qpcrassaytodetectandquantifyshigatoxinproducingecolistecincattleandonfarmsapotentialpredictivetoolforstecculturepositivefarms AT koendereu qpcrassaytodetectandquantifyshigatoxinproducingecolistecincattleandonfarmsapotentialpredictivetoolforstecculturepositivefarms |
_version_ |
1725836716719210496 |