VIEW ARTICLE

Vector Relations

Detection of Rice Ragged Stunt Virus in Insect Vectors by Enzyme-Linked Immunosorbent Assay. Hiroyuki Hibino, Plant Pathologist, Institute for Plant Virus Research, Tsukuba Science City, Yatabe, Ibaraki, 305 Japan; Ikuo Kimura, plant Pathologist, Institute for Plant Virus Research, Tsukuba Science City, Yatabe, Ibaraki, 305 Japan. Phytopathology 72:656-659. Accepted for publication 10 September 1981. Copyright 1982 The American Phytopathological Society. DOI: 10.1094/Phyto-72-656.

Rice ragged stunt virus (RRSV) was purified from diseased rice plants, and antiserum against RRSV was prepared and used for virus detection by enzyme-linked immunosorbent assay (ELISA). RRSV was detected by ELISA in extracts of RRSV-containing rice leaves and newly emerged plant hopper vectors, Nilaparvata lugens, diluted up to 320 and 5,120 times respectively, with phosphate buffer, pH 7.4. However, strong nonspecific reactions occurred in extracts of virus-free female plant hoppers carrying eggs. Polyvinylpyrrolidone reduced the intensity of the nonspecific reaction. The intensity of the nonspecific reaction varied remarkably with pH of the extraction buffer and was lowest at pH 6.5 or 6.0. RRSV was detected in extracts of viruliferous insects diluted up to 10,240 times with phosphate buffer, pH 6.5, containing 2% polyvinylpyrrolidone. Plant hopper extracts in this buffer showed negligible nonspecific reactions. RRSV was detected efficiently from single plant hoppers by ELISA. In tests of plant leafhoppers fed on RRSV-infected plants, 27% transmitted the virus and gave a positive ELISA; 25% did not transmit the virus but gave a positive ELISA; the remainder gave negative ELISA and transmission tests. RRSV was detected in killed insects stored for 1 day at –80 C, 6 C or at room temperature.