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

Karen Verstraete²  Els Van Coillie²  Hadewig Werbrouck²  Stephanie Van Weyenberg²  Lieve Herman²  Jurgen Del-Favero¹  Peter De Rijk¹  Lieven De Zutter³  Maria-Adelheid Joris³  Marc Heyndrickx² 

[1] Applied Molecular Genomics Group, Department of Molecular Genetics, Flemish Institute for Biotechnology (VIB), Universiteitsplein 1, Antwerpen 2610, Belgium; E-Mails:;Technology and Food Science Unit, Institute for Agricultural and Fisheries Research (ILVO), Brusselsesteenweg 370, Melle 9090, Belgium; E-Mails:;Department of Veterinary Public Health and Food Safety, Ghent University, Salisburylaan 133, Merelbeke 9820, Belgium; E-Mails:

关键词: Shiga toxin;    E. coli;    real-time PCR;    cattle;    quantification;    intimin;    screening;    farm;    feces;    isolation;   

DOI  :  10.3390/toxins6041201

来源: mdpi

【 摘 要 】

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.

【 授权许可】

CC BY   
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