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Automation on the Plate Analysis from Yellow Fever Serum Neutralization Assays
Caio B Denani1, Bruno P Setatino2, Renan Oliveira Vieira2
1Laboratory of Immunomolecular Analysis - LANIM, Institute of Technology in Immunobiologicals (Bio-Manguinhos), Oswaldo Cruz Foundation (Fiocruz), Rio de Janeiro, Brazil. caio.denani@bio.fiocruz.br.
Methods in Molecular Biology (Clifton, N.J.)
|April 18, 2025
Summary
Automated systems improve yellow fever virus neutralization assays. These methods enhance accuracy and throughput for vaccine development and public health by minimizing human error in plaque counting.
Area of Science:
- Virology
- Immunology
- Biotechnology
Background:
- Quantifying neutralizing antibodies is essential for antiviral vaccine development and effectiveness evaluation.
- The Plaque-Reduction Neutralization Test (PRNT) is standard but limited by time, throughput, and manual counting errors.
- Existing PRNT variations (FRNT, microFRNT) still suffer from inter-analyst variability due to visual plaque recognition.
Purpose of the Study:
- To describe automated image acquisition and quantification procedures for serum neutralization assays.
- To enhance the throughput and accuracy of neutralizing antibody assays.
- To minimize human inherent variations in plaque/focus counting.
Main Methods:
- Utilizing automated systems integrating microscopy, robotics, and IT for standardized image acquisition and analysis.
- Employing algorithms to identify morphological patterns and quantify structures of interest (e.g., circularity, size, color intensity).
- Implementing automated procedures for plaque/focus counting in neutralization assays.
Main Results:
- Demonstrated increased throughput for serum neutralization assays.
- Achieved improved accuracy in results compared to manual methods.
- Standardized image acquisition and analysis, reducing inter-analyst variability.
Conclusions:
- Automated systems offer a faster and more accurate alternative to traditional methods for quantifying neutralizing antibodies.
- These innovations can significantly aid public health responses to emerging viral diseases.
- Automation minimizes human error, leading to more reliable and reproducible assay results.
