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Please use this identifier to cite or link to this item: http://hdl.handle.net/10373/708

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Title: Quantification of fungal antigens in soil with a monoclonal antibody-based ELISA: analysis and reduction of soil-specific bias
Authors: Otten, Wilfred
Gilligan, Christopher A.
Thornton, C. R.
Affiliation: University of Abertay Dundee. Scottish Informatics, Mathematics, Biology and Statistics Centre
Keywords: Antigen recovery
Freeze-dried mycelium
Issue Date: Jul-1997
Publisher: American Phytopathological Society
Type: Journal Article
Refereed: peer-reviewed
Rights: Published version (c)American Phytopathological Society, available from http://dx.doi.org/10.1094/PHYTO.1997.87.7.730
Citation: Otten, W., Gilligan, C.A. and Thornton, C.R. Quantification of fungal antigens in soil with a monoclonal antibody-based ELISA: analysis and reduction of soil-specific bias. Phytopathology. 87(7): pp.730-736. Available from http://dx.doi.org/10.1094/PHYTO.1997.87.7.730
Abstract: This paper describes methods to improve the use of immunoassays for quantification of soilborne fungal antigens. Calibration curves, prepared by diluting known quantities of an antigen into soil extracts and into soil, were described by a four-parameter logistic curve from which two principal criteria, the lower detection limit and the horizontal locational parameter, were used to summarize the sensitivity and bias of an immuno-assay. We identify two sources of bias, retention of the antigen in soil due to bonding and interference of soluble soil components in plate-trapped—antigen, enzyme-linked immunosorbent assays. Using a monoclonal antibody that recognizes a putative catechol oxidase secreted by hyphae of Rhizoctonia solani, we show that bias due to retention of the antigen in soil is substantially greater than bias due to interference. Three soils were compared: a sand, a clay, and a loam. The degree of retention varied with soil type, with more than a 1,000-fold reduction in sensitivity in the clay soil. Addition of CuSO4 to the extraction solution and optimizing the volume of extractant reduced the bias and increased the sensitivity of the assay for all three soils. Possible mechanisms for the effect due to Cu2+ and the implications for the design and use of calibration curves for assays involving quantification of fungal antigens in soil are discussed.
URI: http://hdl.handle.net/10373/708
ISSN: 0031-949X
Appears in Collections:SIMBIOS Collection
Science Engineering & Technology Collection

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