ISHS Acta Horticulturae 236: V International Symposium on Small Fruit Virus Diseases
DETECTION AND QUANTIFICATION OF BLUEBERRY SHOESTRING VIRUS IN ILLINOIA PEPPERI USING DOT-ELISA AND DOT HYBRIDIZATION |
| Authors: | B.T. Terhune, D.C. Ramsdell, K.L. Klomparens |
| Abstract:
Late instars of Illinoia pepperi (MacG.), the aphid vector of blueberry shoestring virus (BBSSV), were allowed to feed on Parafilm sachets containing 50 ug/ml purified BBSSV for acquisition access periods of between 1 and 96 hours. Aphids were assayed via DAS-ELISA, dot-ELISA (on nitrocellulose or nylon membranes), and dot-hybridization on nitrocellulose membrane. Dot-ELISA blots were reproduced to original size on 4 x 5 inch color transparencies. Transparency and autoradiograph spot intensities were scanned at 550 nm on a Gilford Response II transmission spectrophotometer. Multiple regression analysis of a purified virus dilution series, included on each blot, resulted in polynomial models (Y = bo + b1(log x) + b2(log x)2) with coefficients of determination > 0.75. Virus concentrations in individual aphids (ranging from 1 to 30 ng) were interpolated from models. Dot-ELISA on nitrocellulose and nylon membranes were the most sensitive assays for purified BBSSV, with mean detection limits of 0.94 and 1.37 ng BBSSV, respectively. Dot-ELISA on nylon membrane had the lowest background absorbances for non-viruliferous aphids, and the effect on aphid amendments to purified virus was negligible. | |
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