Department of Medical Microbiology, Malmö University Hospital, Lund University, Malmö, Sweden.
J Gen Virol. 2010 Jul;91(Pt 7):1840-8. doi: 10.1099/vir.0.019349-0. Epub 2010 Feb 24.
This study developed and validated a high-throughput human papillomavirus (HPV) serology method based on Luminex technology, using pseudovirions (PsVs) of eight mucosal HPV types (HPV-6, -11, -16, -18, -31, -45, -52 and -58) and two cutaneous HPV types (HPV-5 and -38) bound to heparin-coated beads. Analysis with neutralizing type-specific monoclonal antibodies against the included HPV types indicated the type specificity of the assay. Analysis of negative-control serum samples from 63 children and 71 middle-aged women with up to one lifetime sexual partner indicated high specificity. Positive-control serum samples from subjects with known HPV DNA status or clinical diagnosis found expected sensitivities for most of the HPV types in 219 European serum samples, but lower than expected in 124 samples from Africa. HPV-45 and -52 did not react as expected with the human serum samples. The PsV-Luminex method was used to determine the HPV-seropositivity-associated relative risk for future cervical cancer using 208 serum samples from a prospective study of 18 814 women followed for 23 years, analysed previously with standard HPV-16 ELISA. The PsV-Luminex method gave similar results to ELISA (kappa=0.77). As expected, HPV seropositivities assayed using the PsV-Luminex method found an increased risk of cervical cancer for HPV-16 [odds ratio (OR)=7.7, 95 % confidence interval (CI)=2.6-23] and HPV-31 (OR=4.1, 95 % CI=1.6-10.8), non-significant tendencies for increased risk for other mucosal HPV types and no risk for the cutaneous HPV types. In summary, multiplexed HPV serology using mammalian-derived PsVs selected for native conformation by binding to heparin-coated beads was validated as a high-throughput HPV serological method for most of the analysed HPV types.
本研究开发并验证了一种基于 Luminex 技术的高通量人乳头瘤病毒（HPV）血清学方法，该方法使用八种黏膜 HPV 型（HPV-6、-11、-16、-18、-31、-45、-52 和-58）和两种皮肤型 HPV（HPV-5 和 -38）的假病毒（PsV）结合到肝素包被的珠上。用包含的 HPV 型的中和型特异性单克隆抗体进行分析表明，该测定法具有型特异性。对来自 63 名儿童和 71 名一生中只有一个性伴侣的中年女性的阴性对照血清样本的分析表明，该方法具有高特异性。来自具有已知 HPV DNA 状态或临床诊断的阳性对照血清样本的 219 份欧洲血清样本的大多数 HPV 型的预期敏感性，但在来自非洲的 124 份样本中低于预期。HPV-45 和 -52 与人类血清样本的反应不如预期。使用来自前瞻性研究的 208 份血清样本，该前瞻性研究对 18814 名女性进行了 23 年的随访，该研究使用 Luminex 法检测了 208 份血清样本，之前曾使用标准 HPV-16 ELISA 进行了分析，该法用于确定未来宫颈癌的 HPV 血清阳性相关相对风险。Luminex 法与 ELISA 具有相似的结果（kappa=0.77）。如预期的那样，使用 PsV-Luminex 法检测的 HPV 血清阳性率发现 HPV-16（OR=7.7，95%CI=2.6-23）和 HPV-31（OR=4.1，95%CI=1.6-10.8）的宫颈癌风险增加，其他黏膜 HPV 型的风险增加趋势不明显，皮肤型 HPV 型则没有风险。总之，使用结合肝素包被珠选择天然构象的哺乳动物衍生 PsV 进行多重 HPV 血清学检测被验证为大多数分析 HPV 型的高通量 HPV 血清学方法。
