Laboratoire de santé publique du Québec, Institut national de santé publique Québec, 20045 Sainte-Marie Road, Sainte-Anne-de-Bellevue, Quebec, Canada.
Sainte-Justine Hospital Research Center, Université de Montréal, 3175 Côte Sainte-Catherine, Montreal, Quebec, Canada.
J Gen Virol. 2021 May;102(5). doi: 10.1099/jgv.0.001610.
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 (=0.90), followed by HPV6 (=0.86) and HPV18 (=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, =0.001] and HPV18 [AUC 0.71 versus 0.57, =0.024]. AUCs for HPV6 and HPV11 were similar between the two assays (0.70 versus 0.71, =0.59, and 0.88 versus 0.96, =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.
血清抗体水平可用于衡量针对人乳头瘤病毒（HPV）的体液免疫应答。我们开发并验证了一种快速、技术简单且相对廉价的多重非竞争 Luminex 基于免疫测定法（ncLIA），用于测量针对四种 HPV 类型的总 IgG 抗体水平。该测定法的固相是 HPV6、11、16 和 18 的病毒样颗粒（VLPs），与肝素包被的珠子结合。使用藻红蛋白缀合的二级多克隆驴抗人 IgG 抗体来定量 HPV 血清抗体与 VLPs 的结合。使用来自加拿大蒙特利尔 Hitch 队列研究的 96 对血清和生殖器样本对 ncLIA 进行了标准化和验证，包括年轻女性（18-24 岁）及其男性性伴侣（18 岁以上）。使用 Pearson 相关系数、接收者操作特征曲线和逻辑回归比较了 ncLIA 和 PPD 实验室验证的 Luminex 免疫测定法的结果。我们的测定法具有良好的内和间分析变异性。ncLIA 和验证测定法之间血清抗体水平的相关性对于 HPV16 和 HPV11 最高（=0.90），其次是 HPV6（=0.86）和 HPV18（=0.67）。ncLIA 比验证测定法更能预测生殖器样本中 HPV DNA 的阳性情况，对于 HPV16（曲线下面积（AUC）0.65 与 0.52，=0.001）和 HPV18（AUC 0.71 与 0.57，=0.024）。对于 HPV6 和 HPV11，两种测定法的 AUC 相似（0.70 与 0.71，=0.59，和 0.88 与 0.96，=0.08）。开发的 ncLIA 可用于测量针对四价疫苗中包含的四种 HPV VLPs 的自然感染或疫苗接种后的总 IgG 抗体反应。
