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P3-157 Salmonella enterica Identification and Serotyping from Cilantro Using a PCR Multiplex for Serotyping

Wednesday, July 31, 2013
Exhibit Hall (Charlotte Convention Center)
Junia Jean-Gilles Beaubrun, U.S. Food and Drug Administration, Laurel, MD
Laura Ewing, U.S. Food and Drug Administration, Laurel, MD
Karen Jarvis, U.S. Food and Drug Administration, Laurel, MD
Kim Dudley, U.S. Food and Drug Administration, Laurel, MD
Gopal Gopinath, U.S. Food and Drug Administration, Laurel, MD
Aparna Jayaram, U.S. Food and Drug Administration, Laurel, MD
Jessica Elmore, U.S. Food and Drug Administration, Laurel, MD
Christopher Grim, U.S. Food and Drug Administration-CFSAN-DVA, Laurel, MD
Martha Lamont, U.S. Department of Agriculture, Manassas, VA
Tim McGrath, U.S. Food and Drug Administration, Rockville, MD
Darcy Hanes, U.S. Food and Drug Administration, Laurel, MD
Introduction: In this study a multiplex PCR serotyping method consisting of two five-plex and a single two-plex PCR reactions was used to screen, identify, and serotype Salmonella spp. from 564 cilantro samples using a single non-selective enrichment step.  The anticipated impact from this study is that it will provide a sensitive and specific method for serotyping Salmonella enterica spp. from various food matrices using a single enrichment step.

Purpose: To find a faster and efficient method to identify and serotype Salmonella enterica from produce. 

Methods: Freshly collected MDP cilantro samples were added to mBPW broth and incubated overnight at 37°C. Overnight enrichments were plated on XLT4 agar a selective media used to isolate Salmonella from the cultures. Suspect Salmonella colonies were passaged overnight on 5% sheep blood agar (BA) and assayed on the Vitek compact 2 to confirm Salmonella colonies. DNA was extracted from 24 h enrichment cultures using the NucliSENS easyMag instrument, and PCR analyses were conducted on the BioRad DNA Engine thermocycler and the Agilent 2100 Bioanalyzer. The BAM method and Premi®Test were used as the gold standards for definitive Salmonella identification and serotyping.

Results: From the 564 cilantro samples collected between July 2011 to October 2012, 58% were positive for S. Newport, 32% for S. Tennessee, 10% for S. Montevideo, and one sample for S. SaintPaul. Serotypic patterns were obtained from the 24 h cilantro enrichment broth cultures. The serotypes were confirmed by Check & Trace “Premi®test” for Salmonella.

Significance: The multiplex PCR method for serotyping the 30 most common clinical relevant Salmonella enterica is inexpensive and provided rapid 24-48 h detection capabilities for Salmonella spp. and will be a useful resource for screening foods for the presence of common serotypes.

Back to: Poster Session 3 - Applied Laboratory Methods, Novel Laboratory Methods, Produce, Risk Assessment
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