Development of a rapid and simple immunochromatographic assay to identify Vibrio parahaemolyticus.

Abstract To rapidly and simply determine whether or not bacterial colonies growing on agar were Vibrio parahaemolyticus , we developed an immunochromatographic assay (VP-ICA) using two different monoclonal antibodies (designated mAb-VP34 and mAb-VP109) against the delta subunit of V. parahaemolyticus -F 0 F 1 ATP synthase. The epitopes recognized by mAb-VP34 and mAb-VP109 were mapped to sequences of eight ( 47 LLTSSFSA 54 ) and six amino acid residues ( 16 FDFAVD 21 ), respectively. An amino acid sequence similarity search of the NCBI database using BLASTP showed that both epitopic amino acid sequences were present together only in V. parahaemolyticus . When 124 V. parahaemolyticus strains and 94 strains of 27 other Vibrio species or 35 non- Vibrio species were tested using the VP-ICA, the VP-ICA identified V. parahaemolyticus with 100% accuracy. The VP-ICA rapidly and simply identified the pathogen directly from a single agar colony within 30 min, indicating that VP-ICA will greatly reduce labor and time required to identify V. parahaemolyticus compared with conventional biochemical tests.