一种用于在硝酸纤维素膜上诊断葡萄卷叶病毒的化学发光免疫分析方法。

A chemiluminescent immunoassay for the diagnosis of grapevine closteroviruses on nitrocellulose membrane.

作者信息

Pollini C P, Giunchedi L, Credi R

机构信息

Institute of Plant Pathology, University of Bologna, Italy.

出版信息

J Virol Methods. 1993 Apr;42(1):107-16. doi: 10.1016/0166-0934(93)90182-q.

DOI:10.1016/0166-0934(93)90182-q

PMID:8320305

Abstract

A dot-immunobinding assay was adapted on enhanced chemiluminescence (DIBA-ECL), which employs luminol, a cyclic diacylhydrazide, as substrate for horseradish peroxidase conjugated with a secondary antibody, for the diagnosis of grapevine closteroviruses I and III. The sensitivity of DIBA-ECL was also compared to other immunoenzymatic methods. DIBA-ECL proved to be at least 16 times more sensitive than the dot-immunobinding assay using chloronaphthol/diaminobenzidine mixture as a substrate, which was at least twice as sensitive as DAS-ELISA, DAS-indirect avidin-biotin complex ELISA, and dot-immunobinding assay, using alkaline phosphatase as enzyme. Optimisation of all parameters involved in the DIBA-ECL procedure and its advantages are discussed.

摘要

一种基于增强化学发光的斑点免疫结合测定法（DIBA-ECL）被应用于葡萄藤褪绿斑驳病毒I和III的诊断，该方法使用鲁米诺（一种环状二酰肼）作为与二抗偶联的辣根过氧化物酶的底物。DIBA-ECL的灵敏度也与其他免疫酶法进行了比较。结果表明，DIBA-ECL的灵敏度至少比使用氯萘酚/二氨基联苯胺混合物作为底物的斑点免疫结合测定法高16倍，而后者的灵敏度至少是双抗夹心ELISA、间接抗生物素蛋白-生物素复合物ELISA以及使用碱性磷酸酶作为酶的斑点免疫结合测定法的两倍。本文还讨论了DIBA-ECL程序中所有参数的优化及其优势。