DEVELOPMENT

MILITARY PERSONNEL ARE AT RISK TO VARIOUS FORMS OF GASTROENTERITIS AS THEY ARE TRANSFERRED TO PARTS OF THE WORLD WHERE CONDITIONS FAVOR CONTAMINATION OF FOOD OR WATER WITH THE CAUSATIVE AGENTS. MANY OF THESE FORMS OF GASTROENTERITIS ARE SELF LIMITING, BUT VIBRIO CHOLERAE IS THE CAUSATIVE AGENT OF A SEVERE FORM OF GASTROENTERITIS WHICH CAN BE FATAL IF UNTREATED. IT IS THE GOAL OF THIS PROJECT TO DEVELOP A HIGHLY SENSITIVE IMMUNOASSAY WITH AN AMPLIFIED INDICATOR SYSTEM FOR DETECTION OF VIBRIO CHOLERAE IN CLINICAL OR ENVIRONMENTAL SAMPLES. HIGHLY SPECIFIC MONOCLONAL ANTIBODIES WILL BE PRODUCED AGAINST THE ORGANISM, AND CONJUGATED OF THESE ANTIBODIES WILL BE PREPARED FOR USE IN THE DETECTION SYSTEM. THE ASSAY WILL BE DEVELOPED IN VACUUM MICROWELL FORMAT. VARIOUS METHODS OF RAPIDLY EXTRCTING AND CONCENTRATING VIBRIOS FROM SAMPLES AND IMMOBILIZING THEM ON THE MICROWELL MEMBRANES WILL BE COMPARED AND OPTIMIZED. CONCENTRATION METHODS EVALUATED WILL INCLUDE IMMUNOAFFINITY, AND IMMOBILIZATION METHODS EVALUATED WILL INCLUDE FILTRATION IMPINGEMENT, ANTIBODY CAPTURE, AND METAL OXIDE IMMOBILIZATION. EXTRACTION/CONCENTRATION AND ENZYME IMMUNOASSAY PROTOCOLS WILL BE EVALUATED USING SEEDED CLINICAL AND ENVIRONMENTAL SAMPLES.