Concordance assessment between a multiplexed competitive Luminex immunoassay, a multiplexed IgG Luminex immunoassay, and a pseudovirion-based neutralization assay for detection of human papillomaviruse types 16 and 18

There are two approved vaccines against anogenital human papillomaviruses (HPV) and a nine-valent vaccine is currently under development. Although there are several assays available to measure antibodies elicited by HPV vaccines, there is currently no global standard for HPV antibody assays. In the current study, antibody responses to HPV16 and HPV18 among young men and women vaccinated with a quadrivalent HPV6/11/16/18 (qHPV) vaccine were assessed using three assays: a competitive Luminex immunoassay (cLIA-4) which measures antibodies directed against a single neutralizing epitope, an immunoglobulin G Luminex immunoassay (IgG-9) which measures both neutralizing and non-neutralizing antibodies, and a pseudovirion-based neutralization assay (PBNA) which functionally measures the full spectra of neutralizing antibodies. To assess HPV16 and HPV18 responses, 648 and 623 serum samples, respectively, were selected from three prior clinical trials of the qHPV vaccine. For each HPV type, the functional relationship between pairs of assay methods was estimated using a linear statistical relationship model and Pearson correlation coefficients. For both HPV16 and HPV18, the agreement between the PBNA and IgG-9 (correlation coefficients of 0.95 and 0.93, respectively) was comparable to the agreement between the cLIA-4 and IgG-9 (correlation coefficients of 0.92 and 0.92, respectively). Of 478 and 399 post-dose 3 samples that tested positive in the cLIA-4, 100% and 98% also tested positive in the IgG-9 and PBNA. The proportion of cLIA-4 seronegative post-dose 3 samples that tested positive in both the IgG-9 and PBNA was 68% (19/28) for HPV16 and 58% (71/122) for HPV18. The data demonstrate the three assays are highly correlated and reflect the measurement of neutralizing antibody. This further verifies that the IgG-9 assay, which is used to assess the immune response to an investigational nine-valent vaccine, is similarly sensitive to the PBNA for the detection of HPV16 and HPV18 neutralizing antibodies.