Development of a Broad-Spectrum Antigen-Capture ELISA Using Combined Anti-p26 Polyclonal and Monoclonal Antibodies for Detection of Equine Infectious Anemia Virus

TL;DR

This study developed a sensitive and specific ELISA test to detect the EIAV capsid protein p26, offering a reliable method for diagnosing and studying equine infectious anemia virus.

Contribution

The novel AC-ELISA combines monoclonal and polyclonal antibodies for broad-spectrum detection of multiple EIAV strains.

Findings

The AC-ELISA detected p26 with a limit of 1.95 ng/mL and showed a linear range up to 60.5 ng/mL.

The assay effectively distinguished EIAV from other equine viruses without cross-reactivity.

It detected diverse EIAV strains, including virulent, attenuated, and international isolates.

Abstract

Equine Infectious Anemia Virus (EIAV) poses significant diagnostic challenges due to its genetic variability and the limitations of conventional nucleic acid detection methods. This study developed an antigen-capture, enzyme-linked immunosorbent assay (AC-ELISA) for the detection and quantification of the EIAV capsid protein p26. The assay utilized a monoclonal antibody (1G11) specific to the p26 protein as the capture antibody and a polyclonal antibody as the detection antibody, forming a highly specific and sensitive detection system. Under optimized conditions, the detection limit of the AC-ELISA was 1.95 ng/mL, with a good linear relationship observed between 1.95 ng/mL and 60.5 ng/mL of p26 protein. Additionally, the AC-ELISA effectively distinguished EIAV from other equine viruses, including equine herpesvirus 1 (EHV-1), equine arteritis virus (EAV), and equine influenza virus (EIV), without cross-reactivity. Importantly, the AC-ELISA demonstrated the ability to detect multiple EIAV strains, including virulent strains, attenuated strains, and strains from other countries, highlighting its broad applicability across diverse EIAV isolates. Compared to western blot and reverse transcriptase assays, the AC-ELISA exhibited higher sensitivity and strong correlation in quantifying the EIAV p26 protein. The assay is simple, rapid, and cost-effective, making it suitable for both laboratory research and clinical applications. It provides a powerful tool for EIAV detection and quantification, supporting future vaccine development and clinical trials.

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