Application of Anti-H Monoclonal Antibodies to Develop Rapid Immunoassay for Vibrio vulnificus

In the US there are about 76 million foodborne illness cases reported every year, in spite of initiatives by federal agencies. This emphasizes the need for the development of novel detection techniques which are sensitive, specific and rapid. A detection technique should be rapid enough to give results same day, allowing companies to release food lots without delay and decreasing storage costs. This kind of screening would also allow immediate measures, if needed, before releasing the lots. In the case of Vibrio vulnificus (V. vulnificus), conventional methods are available to identify and enumerate this pathogen in oysters, but they are labor-intensive and time consuming. To maintain a constant supply of safe oysters, methods need to be developed that are sensitive and rapid. Application of species-specific monoclonal antibody (MAB) can increase the sensitivity and speed of V. vulnificus detection by eliminating enrichment steps, hence the objective of our study was to develop antibody based rapid and sensitive V .vulnificus detection methods. In the first method monoclonal antibodies were utilized in the development of an immunomagnetic separation (IMS) protocol, which was then combined with rt-PCR to develop rapid method which can detect presence of V. vulnificus in oysters within 3 h with a sensitivity of 102 CFU/ml oyster homogenate. We have also used our anti V. vulnificus -H species specific monoclonal antibodies to develop a lateralflow detection device (dipstick) which when combined with a short 5 h enrichment step was successfully able to identify less than 10 CFU/ ml of V. vulnificus from oyster homogenate. Our IMS rt-PCR and dip stick assay could be an answer to seafood industries rapid pathogen detection needs