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一种改进的放射免疫分析法检测T-2毒素

Detection of T-2 toxin by an improved radioimmunoassay.

作者信息

Fontelo P A, Beheler J, Bunner D L, Chu F S

出版信息

Appl Environ Microbiol. 1983 Feb;45(2):640-3. doi: 10.1128/aem.45.2.640-643.1983.

Abstract

T-2 toxin in serum, urine, and saline was analyzed by a modified radioimmunoassay procedure. The specimens were added directly to the assay tubes without extraction steps. The reaction between antibody and ligands was optimal at 1 h. Albumin-coated charcoal was used to separate bound from free radioactivity. Quenching, which occurred with hemolyzed specimens, was corrected by a wet oxidation process with 60% perchloric acid and 30% hydrogen peroxide. The shorter incubation times resulted in an assay that takes less than 6 h to complete. The average affinity constant of the antibody (Km) was 1.75 X 10(10) liters/mol. The sensitivity was 1 ng per assay or 10 ng/ml. Among the other trichothecenes tested, only H-T-2 cross-reacted significantly (10.3%).

摘要

采用改良的放射免疫分析法对血清、尿液和盐溶液中的T-2毒素进行分析。标本无需提取步骤,直接加入测定管中。抗体与配体之间的反应在1小时时最为理想。用白蛋白包被的活性炭分离结合态和游离态放射性。溶血标本出现的淬灭现象通过用60%高氯酸和30%过氧化氢进行湿氧化处理来校正。较短的孵育时间使得该测定法完成时间少于6小时。抗体的平均亲和常数(Km)为1.75×10¹⁰升/摩尔。灵敏度为每次测定1纳克或10纳克/毫升。在测试的其他单端孢霉烯族毒素中,只有H-T-2有显著交叉反应(10.3%)。

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