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RESEARCH ARTICLE
Contents Vol 57(7)

An enzyme-linked immunosorbent assay for the detection of Rathayibacter toxicus, the bacterium involved in annual ryegrass toxicity, in hay

A. M. Masters A B , A. R. Gregory A , R. J. Evans A , J. E. Speijers A and S. S. Sutherland A
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- Author Affiliations

A Department of Agriculture Western Australia, 3 Baron-Hay Court, South Perth, WA 6151, Australia.

B Corresponding author. Email: amasters@agric.wa.gov.au

Australian Journal of Agricultural Research 57(7) 731-742 https://doi.org/10.1071/AR03125
Submitted: 10 June 2003  Accepted: 6 February 2006   Published: 14 July 2006

Abstract

An enzyme-linked immunosorbent assay (ELISA) for Rathayibacter toxicus is described. The development of a monoclonal antibody for a specific antigen from R. toxicus and a polyclonal antibody raised against the same R. toxicus preparation enabled a capture assay format. The assay is specific for a soluble polysaccharide produced by the bacterium and was found to be sensitive enough to detect antigen equivalent to less than one gall per kilogram of hay. The applicability of the assay to samples of pasture or hay is demonstrated. Cost-effective testing of large numbers of samples for the presence of R. toxicus is possible with the ELISA. This will assist stockowners, hay producers, and hay exporters in the management of the risk of annual ryegrass toxicity.

Additional keywords: ELISA, immunoassay.


Acknowledgments

We thank D. Roberts for supplying the field samples, A. Kelly, G. Dawson, and R. McKenzie of the Queensland Department of Primary Industries and Fisheries for providing the ‘uninfected’ hay, L. den Hollander, D. Hare, M. Kalkhoven, D. Galvin, and N. Lowry for their excellent technical assistance during this study, and Dr J. Allen for advice on the manuscript. The ‘Corynebacterium agropyri’ isolate was kindly provided by Dr Ian Riley. Financial support was provided by the Grains Research Development Corporation and the Rural Industries Research and Development Corporation.


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