A Highly Specific Antibody-Based Assay for Nipah Virus AlphaLISA Detection.

Area of Science:

• Virology
• Immunology
• Biotechnology

Background:

• Nipah virus (NiV) is a zoonotic pathogen causing fatal encephalitis with high case fatality rates.
• Current diagnostic methods for NiV suffer from low sensitivity and long turnaround times.
• The NiV surface glycoprotein (G) is crucial for host cell entry and a potential diagnostic target.

Purpose of the Study:

• To develop a novel, sensitive, and rapid diagnostic assay for Nipah virus.
• To establish an Amplified Luminescent Proximity Homogeneous Assay (AlphaLISA) platform utilizing anti-NiV-G monoclonal antibodies.
• To evaluate the performance characteristics of the developed AlphaLISA system for NiV-G protein detection.

Main Methods:

• Screening of high-affinity anti-NiV-G monoclonal antibodies using flow cytometry and single-cell cloning.
• Optimization of the AlphaLISA reaction system, including antibody conjugation and microsphere ratios.
• Construction and validation of the AlphaLISA detection platform for NiV-G protein.

Main Results:

• The developed AlphaLISA assay achieved a detection sensitivity of 0.024 ng/mL for NiV-G protein, significantly higher than conventional ELISA.
• The assay demonstrated good reproducibility (coefficient of variation <9.5%) and specificity against other zoonotic viruses.
• Detection time was less than 30 minutes, with minimal interference from human serum.

Conclusions:

• A novel, highly sensitive, and rapid AlphaLISA-based diagnostic assay for Nipah virus has been successfully developed.
• The assay shows excellent performance characteristics, including sensitivity, specificity, and speed, making it suitable for clinical applications.
• This innovative diagnostic approach holds significant promise for timely and accurate detection of Nipah virus infections.