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ARS Home » Midwest Area » Ames, Iowa » National Animal Disease Center » Food Safety and Enteric Pathogens Research » Research » Publications at this Location » Publication #202746
Title: COMPARISON OF TWO COMMERCIALLY AVAILABLE REAL-TIME PCR FORMATS FOR THE DETECTION OF SALMONELLA AND CAMPYLOBACTER

Author
item PINNEY, R - IOWA STATE UNIVERSITY
item Muraoka, Wayne
item GRIFFITH, R - IOWA STATE UNIVERSITY
item Wesley, Irene
item TRAMPEL, D - IOWA STATE UNIVERSITY

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 10/1/2006
Publication Date: 10/1/2006
Citation: Pinney, R.M., Muraoka, W.T., Griffith, R., Wesley, I.V., Trampel, D.W. 2006. Comparison of two commercially available real-time PCR formats for the detection of Salmonella and Campylobacter [abstract]. Food Safety Consortium 2006 Symposium. October 1-3, 2006, Fayetteville, Arkansas. 2006 CDROM.

Interpretive Summary:

Technical Abstract: Two commercially available real-time PCR instruments (A and B) were compared for their capabilities to detect Salmonella and Campylobacter species in poultry ceca. Using the USDA-FSIS method of isolating Salmonella as a gold standard, the detection sensitivity and specificity of Instrument A was 94.8% and 100%, respectively. Instrument B had a detection sensitivity and specificity of 95.9% and 96.9%, respectively. In addition, post-enrichment quantification of Salmonella in poultry ceca was estimated using instrument A. Salmonella-free cecal contents (10% wt/vol) were spiked to contain 0-106 CFU/ml of Salmonella abaetetuba as determined by viable plate count. Pre-enrichment cecal suspensions spiked with <5 log10 were below the limits of instrument detection. However, post-enrichment suspensions initially spiked with 2 log10 were detected. This may suggest that enrichment of the Salmonella-spiked cecal contents (20 hrs, 35°C) results in a 2.25 log increase. This factor may be used to extrapolate postenrichment values to pre-enrichment titers. For Campylobacter, selective enrichment followed by selective plating resulted in 18.8% positive samples, while Instrument A and B resulted in 51.1% and 59.0%, respectively. This indicates that real-time PCR detection may be more sensitive than conventional culture methods.