IngentaConnect Interlaboratory Validation of a Real-Time PCR 24-Hour Rapid Metho...

Authors: Cheng, Chorng-Ming¹; Van, Khanh T.¹; Lin, Wen²; Ruby, Richard M.¹

Source: Journal of Food Protection®, Volume 72, Number 5, May 2009 , pp. 945-951(7)

Publisher: International Association for Food Protection

Abstract:

The efficacy of a 24-h Salmonella real-time, or quantitative, PCR (qPCR) detection method was assessed through a collaborative effort involving eight Federal and state laboratories. Eleven foods including mashed potatoes, soft cheese, chili powder, chocolate, eggs, sprouts, apple juice, fish, shrimp, ground beef, and ground chicken were tested. For each food, seven blind samples were distributed to each participant for testing. These included six samples equivalently inoculated with 1 to 5 CFU/25 g of various serotypes of Salmonella (Gaminara, Weltevreden, Heidelberg, Senftenberg, Enteritidis, Newport, Typhimurium, and Kentucky for each food) and 10 to 50 CFU/25 g of the competitor Enterobacter cloacae. The seventh sample was inoculated with 10 to 50 CFU/25 g of the competitor, E. cloacae, only. These samples were tested for Salmonella by using four methods in parallel: (i) 24-h qPCR method detecting Salmonella from modified buffered peptone water enrichment medium; (ii) 48-h qPCR method detecting Salmonella from a secondary selective enrichment broth; (iii) modified Bacteriological Analytical Manual method; and (iv) VIDAS, an immunoassay system. The results of the statistical analysis showed there was no significant (P ≥ 0.05) difference between either of the qPCR methods and the modified Bacteriological Analytical Manual method for 10 of 11 foods. For the one exception, sprouts, detection by qPCR required 48 h. Both qPCR methods showed a detection limit of 0.08 to 0.2 CFU/g. These results provide a solid basis for using this 24-h qPCR rapid screening method to detect Salmonella in foods.

Document Type: Research article

Affiliations: 1: U.S. Food and Drug Administration, Office of Regulatory Affairs, Pacific Regional Laboratory Southwest, 19701 Fairchild, Irvine, California 92612 2: U.S. Food and Drug Administration, Office of Regulatory Affairs, Division of Field Science, 5600 Fishers Lane, Room 12-41, Rockville, Maryland 20857, USA

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Interlaboratory Validation of a Real-Time PCR 24-Hour Rapid Method for Detection of Salmonella in Foods