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Abstract
Serum antibody levels can be used to measure the humoral immune response against human papillomaviruses (HPV). We developed and validated a rapid, technically simple and relatively inexpensive multiplex non-competitive Luminex-based immunoassay (ncLIA) to measure total IgG antibody levels against four HPV types. For the assay’s solid phase, virus-like particles (VLPs) of HPV6, 11, 16 and 18 were bound to heparin-coated beads. HPV serum antibody levels binding to the VLPs were quantified using a phycoerithrin-conjugated secondary polyclonal donkey anti-human IgG antibody. Standardization and validation of the ncLIA were performed using 96 paired serum and genital samples from participants in the HITCH cohort study, including young women (aged 18–24 years) and their male sexual partners (aged 18+) in Montreal, Canada. Results from the ncLIA were compared to a validated Luminex immunoassay from PPD laboratories using Pearson’s correlation coefficients, receiver operating characteristic curves and logistic regression. Our assay had good inter- and intra-assay variability. The correlation of serum antibody levels between the ncLIA and validation assay was highest for HPV16 and HPV11 (r=0.90), followed by HPV6 (r=0.86) and HPV18 (r=0.67). The ncLIA was better able to predict HPV DNA positivity in genital samples than the validation assay for HPV16 [area under the curve (AUC) 0.65 versus 0.52, P=0.001] and HPV18 [AUC 0.71 versus 0.57, P=0.024]. AUCs for HPV6 and HPV11 were similar between the two assays (0.70 versus 0.71, P=0.59, and 0.88 versus 0.96, P=0.08, respectively). The developed ncLIA is useful for measuring total IgG antibody response following natural infection or vaccination against four HPV VLPs included in the quadrivalent vaccine.
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- Accepted:
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Funding
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Tier 2 Canada Research Chair in Sexually Transmitted Infection Prevention
- Principle Award Recipient: BurchellAnn N.
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Réseau FRQS SIDA-MI
- Principle Award Recipient: CoutléeFrançois
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Merck & Co.
- Principle Award Recipient: CouillardMichel
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Abbott Laboratories Canada
- Principle Award Recipient: CouillardMichel
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The Ministry of Health and Social Services of Quebec, QC, Canada
- Principle Award Recipient: CouillardMichel
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Canadian Institutes of Health Research
- Principle Award Recipient: WissingMichel D.
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National Institutes of HealthR01AI073889
- Principle Award Recipient: FrancoEduardo L.
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Canadian Institutes of Health ResearchCRN-83320
- Principle Award Recipient: FrancoEduardo L.
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Canadian Institutes of Health ResearchMOP-68893
- Principle Award Recipient: FrancoEduardo L.