H3N2 influenza hemagglutination inhibition method qualification with data driven statistical methods for human...

Area of Science:

• Virology
• Immunology
• Vaccinology

Background:

• Hemagglutination inhibition (HAI) antibody titers are crucial for assessing vaccine-elicited protection against influenza.
• Current HAI assays lack standardization, leading to variability and low sensitivity across different influenza strains.
• Influenza A (H3N2) is a significant circulating subtype, necessitating reliable methods for vaccine research.

Purpose of the Study:

• To develop and qualify a robust HAI assay for precise quantification of antibody titers against influenza A (H3N2) strains.
• To establish a standardized method for measuring correlates of vaccine-mediated protection in human clinical trials.

Main Methods:

• A hemagglutination inhibition (HAI) assay was designed and qualified using A/Texas/71/2017 as a representative H3N2 strain.
• Guinea pig red blood cells and the neuraminidase inhibitor oseltamivir were utilized.
• Rigorous statistical analysis and strain-specific qualification were performed.

Main Results:

• The qualified HAI assay demonstrated high precision, accuracy, linearity, and robustness (GCV <25%).
• The assay showed excellent specificity and sensitivity.
• Successful qualification was achieved for a second H3N2 strain, A/Singapore/INFIMH-16-0019/2016, meeting all criteria.

Conclusions:

• A qualified and adaptable influenza serology method was established for efficient data generation.
• This method enables accurate measurement of HAI titers to define correlates of vaccine-mediated protection.
• The approach minimizes extra testing and development for new influenza strains.