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An automated ELISA using recombinant antigens for serologic diagnosis of B virus infections in macaques
David Katz1, Wei Shi, Irina Patrusheva
1Viral Immunology Center, Georgia State University, Atlanta, GA, USA. dkatz@gsu.edu
Abstract:
B virus (Macacine herpesvirus 1) occurs naturally in macaques and can cause lethal zoonotic infections in humans. Detection of B virus (BV) antibodies in macaques is essential for the development of SPF breeding colonies and for diagnosing infection in macaques that are involved in human exposures. Traditionally, BV infections are monitored for presence of antibodies by ELISA (a screening assay) and western blot analysis (WBA; a confirmatory test). Both tests use lysates of infected cells as antigens. Because WBA often fails to confirm the presence of low-titer serum antibodies detected by ELISA, we examined a recombinant-based ELISA as a potential alternative confirmatory test. We compared a high-throughput ELISA using 384-well plates for simultaneous antibody screening against 4 BV-related, recombinant proteins with the standard ELISA and WBA. The recombinant ELISA results confirmed more ELISA-positive sera than did WBA. The superiority of the recombinant ELISA over WBA was particularly prominent for sera with low (<500 ELISA units) antibody titers. Among low-titer sera, the relative sensitivity of the recombinant ELISA ranged from 36.7% to 45.0% as compared with 3.3% to 10.0% for WBA. In addition, the screening and confirmatory assays can be run simultaneously, providing results more rapidly. We conclude that the recombinant ELISA is an effective replacement for WBA as a confirmatory assay for the evaluation of macaque serum antibodies to BV.
Insights
A new recombinant ELISA test offers a more sensitive and rapid method for detecting B virus (BV) antibodies in macaques. This advanced assay improves upon traditional western blot analysis, especially for low-titer samples, enhancing diagnostic accuracy in primate health surveillance.
Area of Science:
- Veterinary Virology
- Immunodiagnostics
- Primate Health
Background:
- B virus (Macacine herpesvirus 1) poses a significant zoonotic risk from macaques to humans.
- Accurate detection of B virus antibodies in macaques is crucial for managing Specific Pathogen Free (SPF) colonies and responding to human exposures.
- Current diagnostic methods, ELISA and western blot analysis (WBA), have limitations in confirming low-titer antibody detection.
Purpose of the Study:
- To evaluate a novel recombinant-based ELISA as a superior confirmatory assay for B virus antibodies in macaques.
- To compare the diagnostic performance of the recombinant ELISA against standard ELISA and WBA, particularly for low-titer sera.
- To assess the potential for simultaneous screening and confirmation using the recombinant ELISA for faster results.
Main Methods:
- Development and implementation of a high-throughput 384-well ELISA utilizing four recombinant B virus proteins.
- Comparative analysis of the recombinant ELISA against standard ELISA and WBA using macaque sera.
- Evaluation of assay sensitivity and confirmation rates, with a focus on low-titer samples (<500 ELISA units).
Main Results:
- The recombinant ELISA confirmed a higher proportion of ELISA-positive sera compared to WBA.
- The recombinant ELISA demonstrated significantly greater sensitivity for low-titer antibodies, ranging from 36.7% to 45.0%, versus 3.3% to 10.0% for WBA.
- Simultaneous screening and confirmation capabilities of the recombinant ELISA offer accelerated diagnostic turnaround times.
Conclusions:
- The recombinant ELISA is a highly effective replacement for WBA as a confirmatory test for B virus antibodies in macaques.
- This improved diagnostic approach enhances the reliability of macaque health monitoring and reduces zoonotic risk.
- The recombinant ELISA provides a more sensitive, rapid, and efficient method for B virus antibody detection in primate populations.
