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B. Adil,K. M. Shankar,B. T. Naveen Kumar,Rajreddy Patil,Abhiman Ballyaya,K. S. Ramesh,Sathish Rama Poojary,Omkar V. Byadgi,Prabhugouda Siriyappagouder 대한수의학회 2013 Journal of Veterinary Science Vol.14 No.4
A monoclonal antibody-based flow-through immunoassay (FTA) was developed using a nitrocellulose membrane placed on the top of adsorbent pads enclosed in a plastic cassette with a test zone at the center. The FTA could be completed within 10 min. Clear purple dots against a white background indicated the presence of Aphanomyces (A.)invadans. The FTA limit of detection was 7 μg/mL for A. invadans compared to 56 μg/mL for the immunodot. FTA and polymerase chain reaction (PCR) could detect A. invadans in fish tissue homogenates at a 10-11 dilution compared to a 10-8 dilution by immunodot. In fish suffering from natural cases of epizootic ulcerative syndrome (EUS)collected from Mangalore, India, FTA and PCR could detect A. invadans in 100% of the samples compared to 89.04%detected by immunodot. FTA reagents were stable and produced expected results for 4 months when stored at 4∼8oC. This rapid test could serve as simple and cost-effective on-site screening tool to detect A. invadans in fish from EUS outbreak areas and in ports during the shipment of live or frozen fish.