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Development, Validation, and Standardization of Polymerase Chain Reaction-Based Detection of E. coli O157

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A diagnostic polymerase chain reaction assay was developed for the detection of E. coli O157 as the first part of a multicenter validation and standardization project. The assay is based on amplification of sequences of the rfbE O157 gene and includes an internal amplification control. The selectivity of the assay was evaluated against 155 strains, including 32 E. coli O157, 38 E. coli non-O157, and 85 non-E. coli. It was shown to be highly inclusive (100%) and exclusive (100%). The assay has a 100% detection probability of approximately 2 × 103 cells per reaction.

Document Type: Research Article

Affiliations: 1: Justus-Liebig-University, Institute of Veterinary Food Sciences, Frankfurter str 92, 35392 Giessen, Germany 2: DEFRA Central Science Laboratory, Sand Hutton, York YO41 1LZ, United Kingdom 3: IHP, PO Box 9012, 7200 GN22, De Stoven, 7206 AX, Zutphen, The Netherlands 4: Danish Veterinary Institute, 27 Bulowsvej, DK-1790, Copenhagen 1790, Denmark

Publication date: May 1, 2004

More about this publication?
  • The Journal of AOAC INTERNATIONAL publishes refereed papers and reviews in the fields of chemical, biological and toxicological analytical chemistry for the purpose of showcasing the most precise, accurate and sensitive methods for analysis of foods, food additives, supplements and contaminants, cosmetics, drugs, toxins, hazardous substances, pesticides, feeds, fertilizers and the environment available at that point in time. The scope of the Journal includes unpublished original research describing new analytical methods, techniques and applications; improved approaches to sampling, both in the field and the laboratory; better methods of preparing samples for analysis; collaborative studies substantiating the performance of a given method; statistical techniques for evaluating data. The Journal will also publish other articles of general interest to its audience, e.g., technical communications; cautionary notes; comments on techniques, apparatus, and reagents.
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