基于单克隆抗体的抗原捕获 ELISA 和免疫层析试验条快速检测 H9N2 亚型禽流感病毒。

Antigen-capture ELISA and immunochromatographic test strip to detect the H9N2 subtype avian influenza virus rapidly based on monoclonal antibodies.

机构信息

State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, and National Clinical Research Center for Infectious Diseases, the First Affiliated Hospital, School of Medicine, Zhejiang University, 79 Qingchun Road, Hangzhou, 310003, Zhejiang, China.

出版信息

Virol J. 2021 Oct 2;18(1):198. doi: 10.1186/s12985-021-01671-4.

DOI:10.1186/s12985-021-01671-4

PMID:34600550

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8487345/

Abstract

BACKGROUND

The H9N2 subtype of avian influenza virus (AIV) has become the most widespread subtype of AIV among birds in Asia, which threatens the poultry industry and human health. Therefore, it is important to establish methods for the rapid diagnosis and continuous surveillance of H9N2 subtype AIV.

METHODS

In this study, an antigen-capture enzyme-linked immunosorbent assay (AC-ELISA) and a colloidal gold immunochromatographic test (ICT) strip using monoclonal antibodies (MAbs) 3G4 and 2G7 were established to detect H9N2 subtype AIV.

RESULTS

The AC-ELISA method and ICT strip can detect H9N2 subtype AIV quickly, and do not cross-react with other subtype AIVs or other viruses. The detection limit of AC-ELISA was a hemagglutinin (HA) titer of 4 for H9N2 subtype AIV per 100 μl sample, and the limit of detection of the HA protein of AIV H9N2 was 31.5 ng/ml. The ICT strip detection limit was an HA titer of 4 for H9N2 subtype AIV per 100 μl sample. Moreover, both detection methods exhibited good reproducibility and repeatability, with coefficients of variation < 5%. For detection in 200 actual poultry samples, the sensitivities and specificities of AC-ELISA were determined as 93.2% and 98.1%, respectively. The sensitivities and specificities of the ICT strips were determined as 90.9% and 97.4%, respectively.

CONCLUSIONS

The developed AC-ELISA and ICT strips displayed high specificity, sensitivity, and stability, making them suitable for rapid diagnosis and field investigation of H9N2 subtype AIV.

摘要

背景

禽流感病毒（AIV）的 H9N2 亚型已成为亚洲禽类中最广泛流行的 AIV 亚型，这对家禽业和人类健康构成了威胁。因此，建立快速诊断和持续监测 H9N2 亚型 AIV 的方法非常重要。

方法

本研究建立了使用单克隆抗体（MAbs）3G4 和 2G7 的抗原捕获酶联免疫吸附试验（AC-ELISA）和胶体金免疫层析试验（ICT）条，用于检测 H9N2 亚型 AIV。

结果

AC-ELISA 方法和 ICT 条能够快速检测 H9N2 亚型 AIV，并且与其他亚型 AIV 或其他病毒无交叉反应。AC-ELISA 的检测限为每 100 μl 样品中 4 个血凝素（HA）效价的 H9N2 亚型 AIV，AIV H9N2 的 HA 蛋白检测限为 31.5 ng/ml。ICT 条的检测限为每 100 μl 样品中 4 个血凝素（HA）效价的 H9N2 亚型 AIV。此外，两种检测方法均具有良好的重现性和可重复性，变异系数<5%。在对 200 个实际禽样进行检测时，AC-ELISA 的敏感性和特异性分别为 93.2%和 98.1%。ICT 条的敏感性和特异性分别为 90.9%和 97.4%。