Monoclonal Antibody-based Biotin-Avidin ELISA for the Detection of Soybean Mosaic Virus in Soybean Seeds

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Abstract
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 an 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.