Detection of Shiga toxin-producing Escherichia coli in ground beef using the GeneDisc real-time PCR system
- Eastern Regional Research Center, Agricultural Research Service, US Department of Agriculture, Wyndmoor, PA, USA
Escherichia coli O157:H7 and certain non-O157 Shiga toxin-producing Escherichia coli (STEC) serogroups have emerged as important public health threats. The development of methods for rapid and reliable detection of this heterogeneous group of pathogens has been challenging. GeneDisc real-time PCR assays were evaluated for detection of the stx1, stx2, eae, and ehxA genes and a gene that identifies the O157 serogroup followed by a second GeneDisc assay targeting serogroup-specific genes of STEC O26, O45, O91, O103, O111, O113, O121, O145, and O157. The ability to detect the STEC serogroups in ground beef samples artificially inoculated at a level of ca. 2–20 CFU/25 g and subjected to enrichment in mTSB or buffered peptone water (BPW) was similar. Following enrichment, all inoculated ground beef samples showed amplification of the correct set of target genes carried by each strain. Samples inoculated with STEC serogroups O26, O45, O103, O111, O121, O145, and O157 were subjected to immunomagnetic separation (IMS), and isolation was achieved by plating onto Rainbow agar O157. Colonies were confirmed by PCR assays targeting stx1, stx2, eae, and serogroup-specific genes. Thus, this work demonstrated that GeneDisc assays are rapid, sensitive, and reliable and can be used for screening ground beef and potentially other foods for STEC serogroups that are important food-borne pathogens worldwide.
Keywords: GeneDisc, real-time PCR, STEC, virulence genes, O-group, detection, isolation, E. coli O157:H7
Citation: Fratamico PM and Bagi LK (2012) Detection of Shiga toxin-producing Escherichia coli in ground beef using the GeneDisc real-time PCR system. Front. Cell. Inf. Microbio. 2:152. doi: 10.3389/fcimb.2012.00152
Received: 06 September 2012; Paper pending published: 22 September 2012;
Accepted: 15 November 2012; Published online: 20 December 2012.
Edited by: Nora L. Padola
, Universidad Nacional del Centro de la Provincia de Buenos Aires, Argentina
Reviewed by: Paula M. Lucchesi
, Universidad Nacional del Centro de la Provincia de Buenos Aires, Argentina Gerardo Leotta
, Universidad Nacional de La Plata, Argentina
Copyright © 2012 Fratamico and Bagi. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in other forums, provided the original authors and source are credited and subject to any copyright notices concerning any third-party graphics etc.
*Correspondence: Pina M. Fratamico, Eastern Regional Research Center, Agricultural Research Service, US Department of Agriculture, 600 East Mermaid Lane, Wyndmoor, PA 19038, USA. e-mail: firstname.lastname@example.org