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Comparative Study
. 1987 Feb;15(3):233-47.
doi: 10.1016/0166-0934(87)90101-7.

Detection and Identification of Plant Viruses by ELISA Using Crude Sap Extracts and Unfractionated Antisera

Comparative Study

Detection and Identification of Plant Viruses by ELISA Using Crude Sap Extracts and Unfractionated Antisera

W P Mowat et al. J Virol Methods. .

Abstract

A simple and rapid procedure of enzyme immunoassay (PTA-ELISA) was used to detect and identify viruses in individual plants. Virus antigen in crude leaf extracts was adsorbed directly to a solid-phase support, allowed to react with unfractionated antiserum and the antigen-antibody complex detected with a general purpose conjugate of protein A and enzyme. Viral antigens were trapped most effectively by high bonding polystyrene microtitre plates loaded with leaf extracts prepared in carbonate buffer at pH 9.6. With protein A-alkaline phosphatase conjugate and the substrate p-nitrophenyl phosphate as the antibody-detection system, 18 plant viruses in 8 virus groups were detected reliably and nonspecific reactions did not occur. However, when the substrate 3,3',5,-tetramethyl benzidine was used in conjunction with protein A-horseradish peroxidase conjugate, nonspecific reactions were given by leaf extracts from some uninfected or virus-infected plant species. Where less sensitivity is required than is provided by versions of ELISA that rely on antibody-captured antigen, this method provides a simple and rapid means of detecting and identifying viruses in crude sap extracts with the aid of unfractionated antisera.

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