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Simultaneous Quantitation of Antibodies to Neutralizing Epitopes on Virus-Like Particles for Human Papillomavirus Types 6, 11, 16, and 18 by a Multiplexed Luminex Assay

机译:通过多重Luminex分析法同时定量中和人乳头瘤病毒类型6、11、16和18的病毒样颗粒上抗原决定簇的抗体

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摘要

Several different methods have been developed to quantitate neutralizing antibody responses to human papillomaviruses (HPVs), including in vivo neutralization assays, in vitro pseudoneutralization assays, competitive radioimmunoassays (cRIAs), and enzyme-linked immunosorbent assays. However, each of these techniques possesses one or more limitations that preclude testing large numbers of patient sera for use in natural history studies and large vaccine clinical trials. We describe here a new multiplexed assay, by using the Luminex Laboratory MultiAnalyte Profiling (LabMAP3) assay system, that can simultaneously quantitate neutralizing antibodies to human papillomavirus types 6, 11, 16, and 18 in 50 μl of serum. The HPV-Luminex competitive immunoassay measures titers of polyclonal antibodies in serum capable of displacing phycoerythrin-labeled detection monoclonal antibodies binding to conformationally sensitive, neutralizing epitopes on the respective virus-like particles. This competitive Luminex immunoassay was found to be as sensitive, accurate, and precise as the currently used cRIAs. An effective HPV vaccine will most likely require several distinct genotypes to protect against multiple cancer causing papillomaviruses. The HPV-Luminex immunoassay should prove to be a useful tool in simultaneously quantitating antibody immune responses to multiple HPV genotypes for natural history infection studies and for monitoring the efficacy of prospective vaccines.
机译:已经开发了几种不同的方法来定量对人乳头瘤病毒(HPV)的中和抗体反应,包括体内中和测定,体外假中和测定,竞争性放射免疫测定(cRIAs)和酶联免疫吸附测定。但是,这些技术中的每一种都具有一个或多个限制,无法测试用于自然史研究和大型疫苗临床试验的大量患者血清。我们通过使用Luminex实验室多分析物分析(LabMAP3)分析系统在此描述了一种新的多重分析方法,该方法可以同时定量50μl血清中针对人乳头瘤病毒6、11、16和18型的中和抗体。 HPV-Luminex竞争性免疫测定法可测定血清中多克隆抗体的滴度,这些滴度能够取代藻红蛋白标记的检测单克隆抗体,这些抗体与相应病毒样颗粒上的构象敏感的中和表位结合。发现这种竞争性的Luminex免疫测定与当前使用的cRIAs一样灵敏,准确且精确。有效的HPV疫苗很可能需要几种不同的基因型来预防多种癌症引起的乳头瘤病毒。 HPV-Luminex免疫测定应被证明是同时定量针对多种HPV基因型的抗体免疫反应的有用工具,用于自然史感染研究和监测前瞻性疫苗的有效性。

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