使用单克隆抗体通过一步酶联免疫吸附测定法对传染性胰腺坏死病毒进行快速血清分型。

Rapid serotyping of infectious pancreatic necrosis virus by one-step enzyme-linked immunosorbent assay using monoclonal antibodies.

作者信息

Domínguez J, Babín M, Sánchez C, Hedrick R P

机构信息

INIA, Dpto. de Sanidad Animal, Madrid, Spain.

出版信息

J Virol Methods. 1991 Jan;31(1):93-103. doi: 10.1016/0166-0934(91)90147-r.

DOI:10.1016/0166-0934(91)90147-r

PMID:1707894

Abstract

A one-step ELISA has been developed for detection and serotyping of infectious pancreatic necrosis virus (IPNV) in infected cell cultures using monoclonal antibodies (mAb) raised against strains representing the Sp, Ab and VR 299 serotypes of IPNV. This assay uses a serotype-specific mAb as capture and a mAb directed against a common epitope as detector antibody. Avidin-peroxidase was employed for amplification. The assay was performed in 90 min by simultaneous incubation of the samples, the biotin labelled mAb and the avidin-peroxidase, and detected 37 ng/ml of purified virus. Serotyping of 34 isolates from different areas of Europe, Asia and America showed a total concordance with the results obtained by the neutralization assay.

摘要

已开发出一种一步酶联免疫吸附测定法（ELISA），用于在感染的细胞培养物中检测传染性胰腺坏死病毒（IPNV）并进行血清分型，该方法使用针对代表IPNV的Sp、Ab和VR 299血清型的毒株产生的单克隆抗体（mAb）。该检测方法使用血清型特异性单克隆抗体作为捕获抗体，使用针对共同表位的单克隆抗体作为检测抗体。采用抗生物素蛋白-过氧化物酶进行放大。通过同时孵育样品、生物素标记的单克隆抗体和抗生物素蛋白-过氧化物酶，在90分钟内完成该检测，可检测到37 ng/ml的纯化病毒。对来自欧洲、亚洲和美洲不同地区的34株分离株进行血清分型，结果与中和试验所得结果完全一致。