Hendry R M, Herrmann J E
J Virol Methods. 1983 Jan;6(1):9-17. doi: 10.1016/0166-0934(83)90063-0.
A direct solid-phase enzyme-linked immunoassay for rapid detection and typing of influenza virus was developed utilizing antibodies immobilized by covalent linkage to nylon beads. Covalent linkage of antibody to nylon was accomplished by treatment of partially hydrolyzed nylon with glutaraldehyde. For comparison to conventional enzyme-linked immunosorbent assays (ELISA), IgG fractions were adsorbed to polystyrene beads. Influenza type-specific immunoglobulins coupled to nylon beads were used in an enzyme-linked immunoassay to identify influenza A/USSR/77(H1N1), and A/Texas/75 (H3N2). In titrations of viral antigen, antibody coupled to nylon beads detected 1.9 X 10(4) plaque-forming units (PFU) per assay, whereas 2.2 X 10(5) PFU were required in assays utilizing antibody adsorbed to polystyrene beads. Use of fluorogenic or radioactive substrates for alkaline phosphatase-labeled antibodies increased the sensitivity for virus detection 10-fold with this enzyme, but were only slightly more sensitive than chromogenic substrates with peroxidase-labeled antibody.
利用通过共价连接固定在尼龙珠上的抗体，开发了一种用于快速检测流感病毒并进行分型的直接固相酶联免疫测定法。通过用戊二醛处理部分水解的尼龙，实现抗体与尼龙的共价连接。为了与传统的酶联免疫吸附测定法（ELISA）进行比较，将IgG级分吸附到聚苯乙烯珠上。与尼龙珠偶联的流感型特异性免疫球蛋白用于酶联免疫测定，以鉴定甲型流感病毒/苏联/77（H1N1）和甲型流感病毒/得克萨斯/75（H3N2）。在病毒抗原滴定中，与尼龙珠偶联的抗体每次测定可检测到1.9×10⁴个空斑形成单位（PFU），而在使用吸附到聚苯乙烯珠上的抗体的测定中需要2.2×10⁵个PFU。使用荧光或放射性底物检测碱性磷酸酶标记的抗体，该酶对病毒检测的灵敏度提高了10倍，但仅比使用过氧化物酶标记抗体的显色底物略敏感。
