Detection of picogram quantities of potyviruses using a dot blot immunobinding assay.
Highly sensitive direct visual detection of potyviruses was achieved using a dot blot immunobinding assay (DBIA). The small sample volumes required permit the detection of as little as 0.5 pg virus in purified preparations. The binding of rabbit antibodies could be visualized using goat anti-rabbit IgG (GAR) conjugated to alkaline phosphatase, beta-D-galactosidase, glucose oxidase, or horseradish peroxidase and histochemical substrates. The avidin-biotin system was also useful, but somewhat less sensitive than GAR-enzyme conjugates. Detection of potyviruses in an aphid vector was also attempted, but without success due to endogenous aphid enzymes.
D000469:Alkaline Phosphatase; D000818:Animals; D001042:Aphids / Q000382:microbiology; D001360:Avidin; D001710:Biotin; D004797:Enzyme-Linked Immunosorbent Assay; D005949:Glucose Oxidase; D006735:Horseradish Peroxidase; D007124:Immunoenzyme Techniques; D007158:Immunologic Techniques; D010942:Plant Viruses / Q000302:isolation & purification*; D011817:Rabbits; D001616:beta-Galactosidase
D001360:Avidin; D001710:Biotin; D005949:Glucose Oxidase; D006735:Horseradish Peroxidase; D000469:Alkaline Phosphatase; D001616:beta-Galactosidase