DETECTION AND QUANTIFICATION OF BLUEBERRY SHOESTRING VIRUS IN ILLINOIA PEPPERI USING DOT-ELISA AND DOT HYBRIDIZATION

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 = b_o + 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.

V International Symposium on Small Fruit Virus Diseases

B.T. Terhune, D.C. Ramsdell, K.L. Klomparens

https://doi.org/10.17660/ActaHortic.1989.236.15