Multiplexed serologic assay for nine anogenital human papillomavirus types

Area of Science:

• Immunology
• Virology
• Biotechnology

Background:

• Human papillomavirus (HPV) infections are a significant global health concern.
• VLP-based vaccines are crucial for preventing HPV infections.
• Accurate measurement of immune response to HPV vaccines is essential.

Purpose of the Study:

• To develop and validate a multiplexed immunoassay for detecting human IgG antibodies against nine HPV types.
• To assess the assay's performance in measuring antibody responses following natural infection or VLP-based vaccination.

Main Methods:

• Developed a multiplexed immunoassay using Luminex microspheres conjugated with HPV virus-like particle (VLP) antigens.
• Employed a direct binding format with a phycoerythrin-labeled secondary antibody to detect human IgG antibodies.
• Validated assay performance using pooled rhesus macaque serum, assessing specificity, linearity, and precision.

Main Results:

• The assay demonstrated high specificity (>99%) and good linearity (<7% per 10-fold dilution).
• Overall assay precision was consistently below 19% across various assay components and operators.
• Serostatus cutoffs varied (70-100% seropositive) based on the method used, reflecting antibody levels post-vaccination.

Conclusions:

• The developed multiplexed immunoassay is a reliable tool for quantifying IgG antibodies to nine HPV types.
• This assay is valuable for evaluating immune responses to HPV infection and VLP-based vaccines.
• The assay's performance characteristics support its use in clinical and research settings for HPV serology.