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Journal of Clinical Microbiology, December 2008, p. 4018-4022, Vol. 46, No. 12
0095-1137/08/$08.00+0     doi:10.1128/JCM.01229-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Rapid Multiplex PCR and Real-Time TaqMan PCR Assays for Detection of Salmonella enterica and the Highly Virulent Serovars Choleraesuis and Paratyphi C{triangledown} ,{dagger}

David F. Woods,1 F. Jerry Reen,1 Deirdre Gilroy,2 Jim Buckley,3 Jonathan G. Frye,4 and E. Fidelma Boyd5*

Department of Microbiology, UCC, National University of Ireland, Cork, Ireland,1 Department of Biology, Cork Institute of Technology, Cork, Ireland,2 Veterinary Laboratory, Cork County Council, Cork, Ireland,3 Bacterial Epidemiology and Antimicrobial Resistance Research Unit, Agricultural Research Service, U.S. Department of Agriculture, Athens, Georgia,4 Department of Biological Sciences, University of Delaware, Newark, Delaware 197165

Received 29 June 2008/ Returned for modification 11 September 2008/ Accepted 3 October 2008

Salmonella enterica is a human pathogen with over 2,500 serovars characterized. S. enterica serovars Choleraesuis and Paratyphi C are two globally distributed serovars. We have developed a rapid molecular-typing method to detect serovars Choleraesuis and Paratyphi C in food samples by using a comparative-genomics approach to identify regions unique to each serovar from the sequenced genomes. A Salmonella-specific primer pair based on oriC was designed as an internal control to establish accuracy, sensitivity, and reproducibility. Serovar-specific primer sets based on regions of difference between serovars Choleraesuis and Paratyphi C were designed for real-time PCR assays. Three primer sets were used to screen a collection of over 100 Salmonella strains, and both serovars Choleraesuis and Paratyphi C gave unique amplification patterns. To develop the technique for practical use, its sensitivity for detection of Salmonella spp. in a food matrix was determined by spiking experiments. The technique was also adapted for a real-time PCR rapid-detection assay for both serovars Choleraesuis and Paratyphi C that complements the current procedures for Salmonella sp. isolation and serotyping.

* Corresponding author. Mailing address: Department of Biological Sciences, University of Delaware, Newark, DE 19716. Phone: (302) 831-1088. Fax: (302) 831-2281. E-mail: fboyd{at}udel.edu

{triangledown} Published ahead of print on 15 October 2008.

{dagger} Supplemental material for this article may be found at http://jcm.asm.org/.

Journal of Clinical Microbiology, December 2008, p. 4018-4022, Vol. 46, No. 12
0095-1137/08/$08.00+0     doi:10.1128/JCM.01229-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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