Development of immunological techniques for the detection of the potential causative agent of the brown ring disease

Abstract

Current methods for the detection of the potential causative agent of the brown ring disease (BRD) in clams ( Vibrio P1) involve the isolation of the microorganism from the affected clams, and further biochemical identification. However, by using this methodology, the positive isolation of P1 strains from Spanish affected clams has not been yet successful. In the present study, three immunological techniques (slide agglutination test, immunofluorescence technique, and indirect dot-blot immunoenzymatic assay) have been developed and performed for the specific detection and identification of P1 strains compared to other Vibrio strains which routinely are isolated from clams affected by BRD. The dot-blot immunoenzymatic technique with P1 antiserum improves both the specificity and practicability of immune identification of Vibrio P1, and only a very weak cross-reactivity was recorded with some V. pelagius-V. splendidus strains isolated from diseased clams. Cross-reactions only occur at high antiserum concentrations and can be overcome by an adequate dilution of the antiserum.