Institute of Biomedicine, University of Turku, Turku, Finland.
Department of Biotechnology, University of Turku, Turku, Finland.
mSphere. 2019 Sep 11;4(5):e00447-19. doi: 10.1128/mSphere.00447-19.
Serological assays are used to diagnose and characterize host immune responses against microbial pathogens. Microarray technologies facilitate high-throughput immunoassays of antibody detection against multiple pathogens simultaneously. To improve survey of influenza A virus (IAV), influenza B virus (IBV), respiratory syncytial virus (RSV), and adenovirus (AdV) antibody levels, we developed a microarray consisting of IAV H1N1, IAV H1N1pdm09 (vaccine), IAV H3N2, IBV Victoria, IBV Yamagata, RSV, AdV type 5 hexon protein, and control antigens printed on the bottom of a microtiter plate well. Bound IgG antibodies were detected with anti-human IgG-coated photon-upconverting nanoparticles and measured with a photoluminescence imager. The performance of the microarray immunoassay (MAIA) was evaluated with serum samples ( = 576) collected from children ( = 288) at 1 and 2 years of age and tested by standard enzyme immunoassays (EIAs) for antibodies to IAV vaccine and RSV. EIAs and MAIA showed substantial to almost perfect agreement (Cohen's κ, 0.62 to 0.83). Applying MAIA, we found seroprevalences of 55% for IAV H1N1, 54% for IAV vaccine, 30% for IAV H3N2, 24% for IBV Victoria, 25% for IBV Yamagata, 38% for RSV, and 26% for AdV in 1-year-old children ( = 768). By the age of 2 years, IgG seropositivity rates ( = 714) increased to 74% for IAV H1N1, 71% for IAV vaccine, 49% for IAV H3N2, 47% for IBV Yamagata, 49% for IBV Victoria, 68% for RSV, and 58% for AdV. By analyzing increases in antibody levels not biased by vaccinations, we found a reinfection rate of 40% for RSV and 31% for AdV in children between 1 and 2 years of age. The multiplex immunoassay was successfully used to simultaneously detect antibodies against seven different viruses. The developed serological microarray is a new promising tool for diagnostic, epidemiological, and seroprevalence analyses of virus infections.
血清学检测用于诊断和描述宿主对微生物病原体的免疫反应。微阵列技术有助于同时高通量检测针对多种病原体的抗体检测。为了提高对甲型流感病毒（IAV）、乙型流感病毒（IBV）、呼吸道合胞病毒（RSV）和腺病毒（AdV）抗体水平的调查，我们开发了一种微阵列，其中包含 H1N1、H1N1pdm09（疫苗）、H3N2、Victoria、Yamagata、RSV、AdV 型 5 衣壳蛋白和对照抗原，这些抗原印在微量滴定板孔的底部。结合的 IgG 抗体用抗人 IgG 包被的光上转换纳米粒子检测，并通过光致发光成像仪测量。通过标准酶免疫分析（EIAs）对 IAV 疫苗和 RSV 抗体进行检测，用来自 1 岁和 2 岁儿童（分别为 288 人）的血清样本（ = 576）评估微阵列免疫分析（MAIA）的性能。EIA 和 MAIA 显示出实质性到几乎完美的一致性（Cohen's κ，0.62 到 0.83）。应用 MAIA，我们发现 1 岁儿童的 IAV H1N1 血清流行率为 55%，IAV 疫苗为 54%，IAV H3N2 为 30%，IBV Victoria 为 24%，IBV Yamagata 为 25%，RSV 为 38%，AdV 为 26%。到 2 岁时，IgG 血清阳性率（ = 714）增加到 IAV H1N1 为 74%，IAV 疫苗为 71%，IAV H3N2 为 49%，IBV Yamagata 为 47%，IBV Victoria 为 49%，RSV 为 68%，AdV 为 58%。通过分析不受疫苗接种影响的抗体水平的增加，我们发现 1 至 2 岁儿童的 RSV 再感染率为 40%，AdV 为 31%。该多重免疫分析成功用于同时检测七种不同病毒的抗体。开发的血清学微阵列是一种用于病毒感染诊断、流行病学和血清流行率分析的有前途的新工具。
