Fouda Genevieve G, Leke Rose F G, Long Carole, Druilhe Pierre, Zhou Ainong, Taylor Diane Wallace, Johnson Armead H
Department of Biology, Georgetown University, Washington, DC 20057, USA.
Clin Vaccine Immunol. 2006 Dec;13(12):1307-13. doi: 10.1128/CVI.00183-06. Epub 2006 Oct 11.
Antibodies to Plasmodium falciparum are classically measured using the enzyme-linked immunosorbent assay (ELISA). Although highly sensitive, this technique is labor-intensive when large numbers of samples must be screened against multiple antigens. The suspension array technology (SAT) might be an alterative to ELISA, as it allows measurement of antibodies against multiple antigens simultaneously with a small volume of sample. This study sought to adapt the new SAT multiplex system for measuring antibodies against nine malarial vaccine candidate antigens, including recombinant proteins from two variants of merozoite surface protein 1, two variants of apical merozoite antigen 1, erythrocyte binding antigen 175, merozoite surface protein 3, and peptides from the circumsporozoite protein, ring erythrocyte surface antigen, and liver-stage antigen 1. Various concentrations of the antigens were coupled to microspheres with different spectral addresses, and plasma samples from Cameroonian adults were screened by SAT in mono- and multiplex formats and by ELISA. Optimal amounts of protein required to perform the SAT assay were 10- to 100-fold less than that needed for ELISA. Excellent agreement was found between the single and multiplex formats (R > or = 0.96), even when two variants of the same antigen were used. The multiplex assay was rapid, reproducible, required less than 1 mul of plasma, and had a good correlation with ELISA. Thus, SAT provides an important new tool for studying the immune response to malaria rapidly and efficiently in large populations, even when the amount of plasma available is limited, e.g., in studies of neonates or finger-prick blood.
恶性疟原虫抗体的经典检测方法是酶联免疫吸附测定（ELISA）。尽管该技术灵敏度很高，但当需要针对多种抗原对大量样本进行筛查时，它需要耗费大量人力。悬浮阵列技术（SAT）可能是ELISA的一种替代方法，因为它能够使用少量样本同时检测针对多种抗原的抗体。本研究旨在对新的SAT多重检测系统进行改进，以检测针对九种疟疾疫苗候选抗原的抗体，这些抗原包括来自裂殖子表面蛋白1两种变体、顶端裂殖子抗原1两种变体、红细胞结合抗原175、裂殖子表面蛋白3的重组蛋白，以及来自环子孢子蛋白、环状红细胞表面抗原和肝期抗原1的肽段。将不同浓度的抗原与具有不同光谱特征的微球偶联，然后采用SAT单重和多重检测形式以及ELISA对喀麦隆成年人的血浆样本进行筛查。进行SAT检测所需的最佳蛋白量比ELISA所需的蛋白量少10至100倍。即使使用同一抗原的两种变体，单重和多重检测形式之间也具有极佳的一致性（R≥0.96）。多重检测快速、可重复，所需血浆量少于1微升，并且与ELISA具有良好的相关性。因此，即使在可获得的血浆量有限的情况下，例如在新生儿或指尖血研究中，SAT也为在大量人群中快速有效地研究疟疾免疫反应提供了一种重要的新工具。
