Measurement of antibodies to pneumococcal, meningococcal and haemophilus polysaccharides, and tetanus and diphtheria toxoids using a 19-plexed assay
Measurement of antibodies to pneumococcal, meningococcal and haemophilus polysaccharides, and tetanus and diphtheria toxoids using a 19-plexed assay
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DOI：
10.1016/j.jim.2012.01.007
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发表时间：
2012-03-30
影响因子：
2.2
通讯作者：
MacLennan, Calman A.
中科院分区：
文献类型：
--
作者：
Whitelegg, Alison M. E.;Birtwistle, Jane;MacLennan, Calman A.
抗体对疫苗应答的测量在评估疑似免疫缺陷患者的免疫状态时是有用的。以前依赖于酶联免疫吸附测定法（ELISA）繁琐、耗时且昂贵。流式细胞术系统的可用性导致了多重检测的发展，能够同时测量几种抗原的抗体。我们优化了一种基于流式细胞术的检测方法，用于测定血清中含有19种抗原的IgG和IgM浓度，这些抗原包含在细菌亚单位疫苗组中：肺炎球菌疫苗、脑膜炎球菌疫苗、b型流感嗜血杆菌（Hib）、破伤风和白喉类毒素疫苗。89-SF作为标准血清。该测定法用于测定193名健康成人献血者血清中的特异性抗体水平。IgG和肺炎球菌IgM抗体浓度可在3 log10范围内测量，包括每种抗原的阈值保护性IgG抗体水平。与单组化法相比，19组化法测定的抗体之间几乎没有干扰，缺乏交叉反应性IgG抗体，但有证据表明3/19肺炎球菌抗原存在交叉反应性IgM抗体。193份成人血清15/19 IgG和12/12 IgM浓度的90百分位值均在此范围内，血清中含有保护性IgG抗体的百分比根据抗原的不同从4%到95%不等。这种多重检测方法可以同时检测19种细菌疫苗抗原的抗体水平。它适合用于标准的临床实践，以评估体内免疫反应，以测试疫苗接种和测量这些抗原的绝对抗体水平。(C) 2012 Elsevier B.V.版权所有
The measurement of antibody responses to vaccination is useful in the assessment of immune status in suspected immune deficiency. Previous reliance on enzyme-linked immunoabsorbent assays (ELISA) has been cumbersome, time-consuming and expensive. The availability of flow cytometry systems has led to the development of multiplexed assays enabling simultaneous measurement of antibodies to several antigens. We optimized a flow cytometric bead-based assay to measure IgG and IgM concentrations in serum to 19 antigens contained in groups of bacterial subunit vaccines: pneumococcal vaccines, meningococcal vaccines, Haemophilus influenzae b (Hib), and tetanus and diphtheria toxoid vaccines. 89-SF was employed as the standard serum. The assay was used to determine specific antibody levels in serum from 193 healthy adult donors. IgG and pneumococcal IgM antibody concentrations were measurable across 3 log10 ranges encompassing the threshold protective IgG antibody levels for each antigen. There was little interference between antibody measurements by the 19-plexed assay compared with monoplexed assays, and a lack of cross-reactive IgG antibody, but evidence for cross-reacting IgM antibody for 3/19 pneumococcal antigens. 90th centile values for 15/19 IgG concentrations and 12/12 IgM concentrations of the 193 adult sera were within these ranges and percentages of sera containing protective IgG antibody levels varied from 4% to 95% depending on antigen. This multiplexed assay can simultaneously measure antibody levels to 19 bacterial vaccine antigens. It is suitable for use in standard clinical practice to assess the in vivo immune response to test vaccinations and measure absolute antibody levels to these antigens. (C) 2012 Elsevier B.V. All rights reserved.