Development and Validation of a Multiplex Serology Assay for HPV Type-Specific Antibody Detection

Abstract

Human papillomavirus (HPV) is a common viral infection with over 200 identified types, some of which are classified as oncogenic due to their association with various cancers. Among these, HPV16 is notably the most prevalent and is implicated in the majority of cervical cancers, as well as significant proportions of oropharyngeal, anal, penile, vulvar, and vaginal cancers. Prophylactic vaccines, such as the bivalent, quadrivalent, and 9- valent HPV vaccines, are highly effective in preventing infections with the most common oncogenic HPV types. Despite this, vaccine uptake varies globally due to factors such as vaccine availability, cost, cultural acceptance, and lack of awareness. Consequently, many individuals remain unvaccinated and at risk for HPV infection and its associated cancers. Catch-up vaccination programs are essential for reaching those who missed routine vaccination during adolescence. However, identifying individuals who are seronegative for HPV-16 and other oncogenic types is crucial for the efficient allocation of vaccines, especially in resource-limited settings. Current serological assays are often limited in scope and require large sample volumes, making them less practical for widespread use. We aimed to develop a multiplex serology assay that can simultaneously detect antibodies against multiple HPV types from a small sample volume, providing a valuable tool for targeted vaccination strategies. Also, our objective was to test the performance of our HPV type-specific serology assay using an Internationally defined proficiency panel developed at the National Cancer Institute (NCI) Frederick Laboratory