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Aims:  Multicentre evaluation of biochemical and molecular methods for the identification of Vibrio parahaemolyticus. Methods and Results:  For the biochemical identification methods, API 20E and API 20NE and Alsina's scheme were evaluated in intra‐ and interlaboratory tests in order to determine the accuracy and concordance of each method. Both in intra‐ and interlaboratory tests, the Alsina's scheme showed the highest sensitivity (86% of correct identifications in the interlaboratory test). False‐positive results were obtained by all methods (specificity was 95% for API 20E, 73% for API 20NE and 84% for Alsina's scheme) and concordance varied from 65% of API 20NE to 84% of API 20E. For the molecular identifications, polymerase chain reaction (PCR) for the detection of toxR gene, tl gene and pR72H fragment were tested on 30 strains by two laboratories. The PCR for toxR showed the highest inclusivity (96%), exclusivity (100%) and concordance (97%). Conclusions:  Among the biochemical identification methods tested, the Alsina's scheme gave more reliable results; however, in order to avoid false‐positive results, all the biochemical identifications should be confirmed by means of molecular methods. Significance and Impact of the Study:  Availability of an efficient identification method of Vibrio parahaemolyticus to use in official control of fisheries products.

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