A direct monoclonal antibody sandwich immunoassay for detection of Bonamia ostreae (Ascetospora) in hemolymph samples of the flat oyster Ostrea edulis (Mollusca Bivalvia)

Abstract

Currently recognltlon of molluscan dlseases relles heavlly upon llght mlcroscoplc examlnatlon of stained histological sections of molluscan tissues Such histopathological methods are time- consuming and relatively inefficient In the study of ongolng epizootics Alternatively, more effluent investigative tools are needed to accelerate d ~s e a s e recognltlon and quantif~catlon Such methods are needed to prevent control and eradicate molluscan dlseases Thls paper reports the development of a direct sandwich enzyme-linked lmmunosorbent assay (ELISA) for detecting the ascetosporan intrahemocytic parasite B o n a m ~a ostreae of the flat oyster Ostrea eduhs Bonamiasls 1s a commercially important epizootic dlsease of oysters A Bonam~a-speciflc monoclonal antlbody IS employed in the assay The assay is performed In antibody-coated microplates employng standardized reagent solutions and 50 ul oyster hemolymph samples This procedure was developed into a commercial product, available as a kit Based upon cllnical studles, there was a 90 O/o agreement In test results when ELISAs were compared to the reference standard, hlstopathologic light microscopic examlnatlon for the parasites O Inter-Research/Pnnted ln Germany