使用斑点印迹免疫结合试验检测皮克级数量的马铃薯Y病毒属病毒。

Detection of picogram quantities of potyviruses using a dot blot immunobinding assay.

作者信息

Berger P H, Thornbury D W, Pirone T P

出版信息

J Virol Methods. 1985 Oct;12(1-2):31-9. doi: 10.1016/0166-0934(85)90005-9.

Abstract

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.

摘要

使用斑点印迹免疫结合测定法（DBIA）实现了对马铃薯Y病毒的高灵敏度直接视觉检测。所需的小样本量能够检测纯化制剂中低至0.5 pg的病毒。兔抗体的结合可以使用与碱性磷酸酶、β-D-半乳糖苷酶、葡萄糖氧化酶或辣根过氧化物酶及组织化学底物偶联的山羊抗兔IgG（GAR）来可视化。抗生物素蛋白-生物素系统也很有用，但灵敏度略低于GAR-酶偶联物。还尝试检测蚜虫载体中的马铃薯Y病毒，但由于蚜虫内源性酶而未成功。