The Detection of Low Concentrations of Double-Stranded Ribonucleic Acid with Iodine-125 Labeled Antiserum

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Abstract
Antiserum prepared against double-stranded (ds) RNA poly I .cntdot. poly C reacted in Ouchterlony double-diffusion tests with poly I .cntdot. poly C (titer = 1:16) and poly A .cntdot. poly U (titer = 1.8), but not with yeast RNA or calf thymus DNA. The antiserum also removed the poly I .cntdot. poly C zone from centrifuged sucrose density gradients. When the antiserum against poly I .cntdot. poly C was labeled with 125I and added to varying concentrations of poly I .cntdot. poly C, it detected as little as 10 ng in 0.1 ml of buffer. This labeled antiserum also reacted with similar concentrations of pea enation mosaic virus (PEMV) replicative form (RF) RNA, .vphi. RNA and poly A .cntdot. poly U. Labeled normal serum gave only background counts when tested with any of these RNA. The labeled antiserum against poly I .cntdot. poly C reacted with dsRNA in nucleic acids extracted from nuclei or vesicles of PEMV-infected [pea] cells, but not with nuclei acids extracted from chloroplasts of PEMV-infected cells or with nucleic acids extracted from any healthy cell fraction.

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