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Applied and Environmental Microbiology, December 2000, p. 5472-5476, Vol. 66, No. 12
0099-2240/00/$04.00+0

Simultaneous Detection of Salmonella Strains and Escherichia coli O157:H7 with Fluorogenic PCR and Single-Enrichment-Broth Culture

Vijay K. Sharma* and Steve A. Carlson

Pre-Harvest Food Safety and Enteric Diseases Research Unit, National Animal Disease Center, Agricultural Research Service, USDA, Ames, Iowa 50010

Received 3 July 2000/Accepted 11 September 2000

A multiplex fluorogenic PCR assay for simultaneous detection of pathogenic Salmonella strains and Escherichia coli O157:H7 was developed and evaluated for use in detecting very low levels of these pathogens in meat and feces. Two sets of primers were used to amplify a junctional segment of virulence genes sipB and sipC of Salmonella and an intragenic segment of gene eae of E. coli O157:H7. Fluorogenic reporter probes were included in the PCR assay for automated and specific detection of amplified products. The assay could detect <10 CFU of Salmonella enterica serovar Typhimurium or E. coli O157:H7 per g of meat or feces artificially inoculated with these pathogens and cultured for 6 to 18 h in a single enrichment broth. Detection of amplification products could be completed in <= 4 h after enrichment.

* Corresponding author. Mailing address: USDA, ARS, National Animal Disease Center, P.O. Box 70, Ames, IA 50010. Phone: (515) 663-7406/7279. Fax: (515) 663-7458. E-mail: vsharma{at}nadc.ars.usda.gov.

Applied and Environmental Microbiology, December 2000, p. 5472-5476, Vol. 66, No. 12
0099-2240/00/$04.00+0


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