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Applied and Environmental Microbiology, February 2001, p. 721-724, Vol. 67, No. 2
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.2.721-724.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Evaluation of Two Direct Plating Methods Using Nonradioactive Probes for Enumeration of Vibrio parahaemolyticus in Oysters

J. A. Gooch,1,* A. DePaola,2 C. A. Kaysner,3 and D. L. Marshall4

U. S. Department of Commerce NOAA NOS Center for Coastal Environmental Health and Biomolecular Research, Charleston, South Carolina 29412-91101; U.S. FDA Gulf Coast Seafood Laboratory, Dauphin Island, Alabama 36528-01582; U. S. FDA Seafood Products Research Center, Bothell, Washington 98021-44213; and Department of Food Science and Technology, Mississippi State University, Mississippi State, Mississippi 39762-98054

Received 22 August 2000/Accepted 23 November 2000

Oysters (Crassostrea virginica) were collected monthly from May 1998 to April 1999 from Mobile Bay, Ala., and analyzed to determine Vibrio parahaemolyticus densities at zero time and after 5, 10, and 24 h of postharvest storage at 26°C. After 24 h of storage at 26°C, oysters were transferred to a refrigerator at 3°C and then analyzed 14 to 17 days later. The V. parahaemolyticus numbers were determined by the most-probable-number procedure using alkaline phosphatase-labeled DNA probe VPAP, which targets the species-specific thermolabile hemolysin gene (tlh), to identify suspect isolates (MPN-VPAP procedure). Two direct plating methods, one using a VPAP probe (Direct-VPAP) and one using a digoxigenin-labeled probe (Direct-VPDig) to identify suspect colonies, were compared to the MPN-VPAP procedure. The results of the Direct-VPAP and Direct-VPDig techniques were highly correlated (r = 0.91), as were the results of the Direct-VPAP and MPN-VPAP procedures (r = 0.91). The correlation between the Direct-VPDig and MPN-VPAP results was 0.85. The two direct plating methods in which nonradioactive DNA probes were used were equivalent to the MPN-VPAP procedure for identification of total V. parahaemolyticus, and they were more rapid and less labor-intensive.

* Corresponding author. Mailing address: U. S. Department of Commerce NOAA NOS Center for Coastal Environmental Health and Biomolecular Research, 219 Ft. Johnson Road, Charleston, SC 29412-9110. Phone: (843) 762-8643. Fax: (843) 762-8700. E-mail: jan.gooch{at}noaa.gov.

Applied and Environmental Microbiology, February 2001, p. 721-724, Vol. 67, No. 2
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.2.721-724.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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Copyright © 2001 by the American Society for Microbiology. All rights reserved.