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Disease Detection and Losses

Preparation and Characterization of Monoclonal Antibodies to Double-Stranded RNA. C. A. Powell, University of Florida, IFAS, Agricultural Research and Education Center, P.O. Box 248, Fort Pierce, FL 34954; Phytopathology 81:184-187. Accepted for publication 10 September 1990. Copyright 1991 The American Phytopathological Society. DOI: 10.1094/Phyto-81-184.

Monoclonal antibodies to double-stranded RNA were prepared by fusing splenocytes from BALB/c mice, which had been immunized with poly A · poly U and poly I · poly C, with NS-1 mouse myeloma cells. Of 103 independently derived hybridoma lines which secreted antibodies that reacted with dsRNA, only two competed favorably with rabbit polyclonal antiserum to poly A · poly U for sites on this synthetic RNA. Further characterization of these two monoclonal antibodies showed that they could detect 3 ng poly A · poly U or 10 ng poly I · poly C/ml in indirect double antibody sandwich enzyme-linked immunosorbent assay tests. The antibodies did not react with double-stranded DNA, rRNA, or tRNA. They could distinguish between total nucleic extracts from virus-infected and uninfected plants in some but not all cases.