Serological assays detecting antibodies in serum or plasma samples are useful and versatile instruments to investigate an individual's infection and vaccination history, e.g. for clinical diagnosis, personal risk evaluation, and seroepidemiological studies. Multiplex Serology is a suspension bead array-based high-throughput methodology for simultaneous measurement of antibodies against multiple pathogens in a single reaction vessel, thus economizing sample volume, measurement time, and costs. We developed and validated bead-based pathogen-specific Monoplex Serology assays, i.e. assays including only antigens for the respective pathogen, to detect antibodies against Corynebacterium diphtheriae and Clostridium tetani toxins, rubella virus and parvovirus B19. The developed assays expand the portfolio of existing pathogen-specific bead-based serology assays and can be efficiently incorporated into larger Multiplex Serology panels. The newly developed Monoplex Serology assays consist of only one antigen per infectious agent, expressed as Glutathione S-transferase-fusion proteins in E. coli. Specificity, sensitivity and Cohen's kappa statistics in comparison with routine clinical diagnostic assays were calculated for serum dilutions 1:100 and 1:1000. All pathogen-specific assays were successfully validated at both serum dilutions with the exception of rubella Monoplex Serology which showed impaired sensitivity (57.6%) at dilution 1:1000. Specificities of successfully validated Monoplex Serology assays ranged from 85.6% to 100.0% (median: 91.7%), and sensitivities from 81.3% to 95.8% (median: 90.9%); agreement with the reference assays ranged from substantial to almost perfect (kappa: 0.66-0.86, median: 0.78). Statistical performance and slim assay design enable efficient incorporation of the developed assays into Multiplex Serology.

中文翻译：

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验证检测针对白喉棒状杆菌和破伤风梭菌毒素、风疹病毒和细小病毒 B19 的抗体以纳入多重血清学

检测血清或血浆样品中抗体的血清学测定是调查个人感染和疫苗接种史的有用且通用的工具，例如用于临床诊断、个人风险评估和血清流行病学研究。多重血清学是一种基于悬浮珠阵列的高通量方法，用于在单个反应容器中同时测量针对多种病原体的抗体，从而节省样本量、测量时间和成本。我们开发并验证了基于微珠的病原体特异性 Monoplex 血清学检测，即仅包括相应病原体抗原的检测，以检测针对白喉棒状杆菌和破伤风梭菌毒素、风疹病毒和细小病毒 B19 的抗体。开发的检测扩展了现有的病原体特异性基于珠子的血清学检测的产品组合，并且可以有效地整合到更大的多重血清学面板中。新开发的 Monoplex 血清学检测由每个传染原仅一种抗原组成，在大肠杆菌中表达为谷胱甘肽 S 转移酶融合蛋白。针对 1:100 和 1:1000 的血清稀释度计算了与常规临床诊断分析相比的特异性、敏感性和 Cohen's kappa 统计数据。除了风疹 Monoplex 血清学在 1:1000 稀释时敏感性受损 (57.6%) 外，所有病原体特异性检测均在两种血清稀释度下均得到成功验证。成功验证的 Monoplex 血清学检测的特异性范围为 85.6% 至 100.0%（中位数：91.7%），灵敏度为 81.3% 至 95.8%（中位数：90.9%）；与参考测定的一致性范围从基本到几乎完美（kappa：0.66-0.86，中值：0.78）。统计性能和纤薄的检测设计能够将开发的检测有效地整合到多重血清学中。