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KMID : 0893420130140040413
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Journal of Veterinary Science
2013 Volume.14 No. 4 p.413 ~ p.419
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Development and standardization of a monoclonal antibody-based rapid flow-through immunoassay for the detection of Aphanomyces invadans in the field
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Adil B
Shankar K. M.
Kumar B. T. Naveen
Patil Rajreddy
Ballyaya Abhiman
Ramesh K. S.
Poojary Sathish Rama
Byadgi Omkar V.
Siriyappagouder Prabhugouda
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Abstract
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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 4~8℃. 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.
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KEYWORD
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diagnosis, epizootic ulcerative syndrome, fish, flow-through, monoclonal antibody
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