Development and evaluation of a new non-competitive Luminex immunoassay detecting antibodies against human papillomavirus types 6, 11, 16 and 18.
Development and evaluation of a new non-competitive Luminex immunoassay detecting antibodies against human papillomavirus types 6, 11, 16 and 18.
复制标题
开发和评估新型非竞争性 Luminex 免疫测定法，用于检测人乳头瘤病毒 6、11、16 和 18 型抗体。
DOI：
10.1099/jgv.0.001610
复制
发表时间：
2021
期刊：
影响因子：
--
通讯作者：
Couillard,Michel
中科院分区：
文献类型：
--
作者：
Trevisan,Andrea;Wissing,MichelD;Dagenais,Carole;Forest,Pierre;Ramanakumar,AgnihotramV;Burchell,AnnN;Franco,EduardoL;Coutlée,François;Couillard,Michel
血清抗体水平可用于测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、技术简单且相对便宜的多重非竞争性基于Luminex的免疫测定法（ncLIA），用于测量针对四种HPV类型的总IgG抗体水平。对于测定的固相，将HPV 6、11、16和18的病毒样颗粒（VLP）结合至肝素包被的珠。使用藻红蛋白缀合的第二多克隆驴抗人IgG抗体定量与VLP结合的HPV血清抗体水平。使用来自HITCH队列研究参与者的96对血清和生殖器样本进行ncLIA的标准化和验证，包括加拿大蒙特利尔的年轻女性（18-24岁）及其男性性伴侣（18岁以上）。使用Pearson相关系数、受试者工作特征曲线和逻辑回归，将ncLIA的结果与PPD实验室经验证的Luminex免疫测定结果进行比较。我们的测定具有良好的测定间和测定内变异性。ncLIA和验证试验之间血清抗体水平的相关性最高的是HPV 16和HPV 11（r=0.90），其次是HPV 6（r=0.86）和HPV 18（r=0.67）。ncLIA对生殖器样本中HPV DNA阳性的预测能力优于HPV 16 [曲线下面积（AUC）0.65 vs 0.52，P =0.001]和HPV 18 [AUC 0.71 vs 0.57，P =0.024]的验证检测。两种方法检测HPV 6和HPV 11的AUC相似（分别为0.70和0.71，P =0.59; 0.88和0.96，P =0.08）。开发的ncLIA可用于测量自然感染或接种四价疫苗中包含的四种HPV VLP后的总IgG抗体应答。
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.