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The kinetics of infection of barley protoplasts with barley stripe mosaic virus (BSMV) was followed by enzyme-linked immunosorbent assay (ELISA) using the double-sandwich procedure. Infected protoplasts were identified by an immunoperoxidase method taking precautions to inhibit endogenous peroxidase. About 70% of the protoplasts were infected with BSMV when the inoculum contained 50 µg/ml purified BSMV in 10 mm-potassium citrate buffer pH 4.7 containing 0.65 m-mannitol and 5 µg/ml poly-l-ornithine. It was estimated by ELISA that about 1400 to 1600 virus particles were adsorbed per protoplast. Following an apparent latent period, virus increased linearly between about 14 and 48 h after inoculation. After 48 h the virus concentration reached between 1.62 × 106 and 2.0 × 106 particles per protoplast.