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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.

Authors
  • Adil, B1
  • Shankar, K M
  • Kumar, B T Naveen
  • Patil, Rajreddy
  • Ballyaya, Abhiman
  • Ramesh, K S
  • Poojary, Sathish Rama
  • Byadgi, Omkar V
  • Siriyappagouder, Prabhugouda
  • 1 Aquatic Health Management Laboratory, Department of Aquaculture, College of Fisheries, Karnataka Veterinary, Animal and Fisheries Sciences University, Mangalore 575002, India. , (India)
Type
Published Article
Journal
Journal of veterinary science
Publication Date
Jan 01, 2013
Volume
14
Issue
4
Pages
413–419
Identifiers
PMID: 23820211
Source
Medline
Keywords
License
Unknown

Abstract

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°C. 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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