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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

(2014) TOXINS. 6(4). p.1201-1221
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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
feces, farm, isolation, quantification, Shiga toxin, REAL-TIME PCR, ENTEROHEMORRHAGIC ESCHERICHIA-COLI, POLYMERASE-CHAIN-REACTION, ANTIGEN GENE-CLUSTER, RAPID DETECTION, MULTIPLEX-PCR, BOVINE FECES, LISTERIA-MONOCYTOGENES, VIRULENCE FACTORS, STOOL SPECIMENS, cattle, screening, intimin, E. coli, real-time PCR

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Chicago
Verstraete, Karen, Els Van Coillie, Hadewig Werbrouck, Stephanie Van Weyenberg, Lieve Herman, Jurgen Del-Favero, Peter De Rijk, et al. 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 (4): 1201–1221.
APA
Verstraete, Karen, Van Coillie, E., Werbrouck, H., Van Weyenberg, S., Herman, L., Del-Favero, J., De Rijk, P., et al. (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(4), 1201–1221.
Vancouver
1.
Verstraete K, Van Coillie E, Werbrouck H, Van Weyenberg S, Herman L, Del-Favero J, et al. 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):1201–21.
MLA
Verstraete, Karen et al. “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 6.4 (2014): 1201–1221. Print.
@article{4420807,
  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.},
  author       = {Verstraete, Karen and Van Coillie, Els and Werbrouck, Hadewig and Van Weyenberg, Stephanie and Herman, Lieve and Del-Favero, Jurgen and De Rijk, Peter and De Zutter, Lieven and Joris, Adelheid and Heyndrickx, Marc and De Reu, Koen},
  issn         = {2072-6651},
  journal      = {TOXINS},
  keywords     = {feces,farm,isolation,quantification,Shiga toxin,REAL-TIME PCR,ENTEROHEMORRHAGIC ESCHERICHIA-COLI,POLYMERASE-CHAIN-REACTION,ANTIGEN GENE-CLUSTER,RAPID DETECTION,MULTIPLEX-PCR,BOVINE FECES,LISTERIA-MONOCYTOGENES,VIRULENCE FACTORS,STOOL SPECIMENS,cattle,screening,intimin,E. coli,real-time PCR},
  language     = {eng},
  number       = {4},
  pages        = {1201--1221},
  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},
  url          = {http://dx.doi.org/10.3390/toxins6041201},
  volume       = {6},
  year         = {2014},
}

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