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Toxins 2014, 6(4), 1201-1221; doi:10.3390/toxins6041201
Article

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

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Received: 3 January 2014; in revised form: 3 March 2014 / Accepted: 11 March 2014 / Published: 27 March 2014
(This article belongs to the Section Bacterial Toxins)
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Abstract: 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.
Keywords: Shiga toxin; E. coli; real-time PCR; cattle; quantification; intimin; screening; farm; feces; isolation Shiga toxin; E. coli; real-time PCR; cattle; quantification; intimin; screening; farm; feces; isolation
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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MDPI and ACS Style

Verstraete, K.; Van Coillie, E.; Werbrouck, H.; Van Weyenberg, S.; Herman, L.; Del-Favero, J.; De Rijk, P.; De Zutter, L.; Joris, M.-A.; Heyndrickx, M.; DeReu, K. 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 2014, 6, 1201-1221.

AMA Style

Verstraete K, Van Coillie E, Werbrouck H, Van Weyenberg S, Herman L, Del-Favero J, De Rijk P, De Zutter L, Joris M-A, Heyndrickx M, DeReu K. 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. 2014; 6(4):1201-1221.

Chicago/Turabian Style

Verstraete, Karen; Van Coillie, Els; Werbrouck, Hadewig; Van Weyenberg, Stephanie; Herman, Lieve; Del-Favero, Jurgen; De Rijk, Peter; De Zutter, Lieven; Joris, Maria-Adelheid; Heyndrickx, Marc; DeReu, Koen. 2014. "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 6, no. 4: 1201-1221.



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