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P3-43 FERN Multi-laboratory Validation of the BAX qPCR Vibrio Assay for Identification of Vibrio Isolates

Wednesday, July 31, 2013
Exhibit Hall (Charlotte Convention Center)
Ruiqing Pamboukian, U.S. Food and Drug Administration, Rockville, MD
Willis Fedio, New Mexico State University, Las Cruces, NM
Jessica Jones, U.S. Food and Drug Administration, Dauphin Island, AL
Paul Browning, New Mexico State University, Las Cruces, NM
John Bowers, U.S. Food and Drug Administration, College Park, MD
FERN Laboratory Cadre, U.S. Food and Drug Administration, Rockville, MD
Angelo DePaola, U.S. Food and Drug Administration, Dauphin Island, AL
Introduction: Seafood related vibrio illnesses are on the rise in the U.S. and abroad. FDA is in the process of validating various real-time PCR assays for detection and quantification of vibrios in seafood. The first phase in this process is to demonstrate specificity of this assay for the intended target organisms using pure cultures. The BAX Vibrio Assay is a multiplex real-time PCR that includes proprietary targets and reagents for simultaneous identification of total V.  cholerae, V. parahaemolyticus and V. vulnificus.

Purpose: To evaluate the BAX Vibrio assay for use in identification and confirmation of V. cholerae, V. parahaemolyticus and V. vulnificus isolates by comparing the sensitivity and specificity of the testing method with the BAM method.

Methods: The BAX Vibrio Assay was compared to the reference method (API 20E, BAM Chapter 9) using a paired-study design for identification of V. cholerae, V. parahaemolyticus and V. vulnificus isolates. Each of the six laboratories analyzed 70 randomly coded isolates including 20 V. vulnificus, 20 V. parahaemolyticus, 20 V. cholerae and 10 isolates of other Vibrio spp. or other bacterial genera.

Results: There was no significant difference (P ≥ 0.05) between the BAX Vibrio Assay and the selected reference method (API 20E) for identification of isolates. The sensitivity of the BAX Vibrio Assay was 98%, 98% and 97% for V. cholerae, V. parahaemolyticus and V. vulnificus, respectively.  The specificity of the BAX Vibrio Assay was 96%, 98%, and 97% for V. cholerae, V. parahaemolyticus and V. vulnificus, respectively. 

Significance: The BAX Vibrio Assay is a reliable and rapid alternative to API 20E for identification of V. cholerae, V. vulnificus, and V. parahaemolyticus isolates. This validation will advantage laboratories by offering an additional reference method for identification of these pathogenic vibrios, including during the next validation phase for detection of these organisms in seafood.

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