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J Gen Virol 66 (1985), 2089-2094; DOI 10.1099/0022-1317-66-10-2089
© 1985 Society for General Microbiology
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Monoclonal Antibody-based Biotin-Avidin ELISA for the Detection of Soybean Mosaic Virus in Soybean Seeds

R. Diaco1, J. H. Hill2, E. K. Hill3, H. Tachibana2 and D. P. Durand1

1 Department of Microbiology
and2 Departments of Plant Pathology, Seed and Weed Sciences, Iowa State University, Ames, Iowa 50011
and3 Marion Laboratories, Kansas City, Missouri 64114, U.S.A.

Two monoclonal antibodies (M-Ab) specific for different epitopes on particles of soybean mosaic virus (SMV) were used in a double-antibody sandwich ELISA (M-Ab ELISA). The non-isotopic immunoassay, which used a biotinylated second antibody and an avidin-alkaline phosphatase detection system to detect SMV in soybean seed extracts, was compared with a polyclonal antibody-based solid-phase radioimmunoassay (SPRIA). M-Ab ELISA detected less than 10 ng of SMV/ml and was more sensitive than the SPRIA which detected 25 ng SMV/ml. Furthermore, M-Ab ELISA required less than 36 h for seed sample assays and only 5.5 h for assays involving purified virus, whereas SPRIA required 3 or 4 days. When seeds from 33 field plots, in which 0%, 30% and 50% of the soybean plants had been inoculated with SMV, were assayed by both systems, results of the two tests correlated for 31 of 33 (94%) seed samples. This suggests that dual-site biotin-avidin M-Ab ELISA systems have potential utility for routine screening of seed samples.

Keywords: SMV, biotin-avidin, ELISA, seeds

Received 21 March 1985; accepted 25 June 1985.




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Copyright © 1985 by the Society for General Microbiology.