Bårnes Guro K, Kristiansen Paul A, Caugant Dominique A, Næss Lisbeth M
Department of Bacteriology and Immunology, Norwegian Institute of Public Health, Oslo, Norway Department of International Health, University of Oslo, Oslo, Norway.
Department of Bacteriology and Immunology, Norwegian Institute of Public Health, Oslo, Norway.
Clin Vaccine Immunol. 2015 Jul;22(7):697-705. doi: 10.1128/CVI.00087-15. Epub 2015 Apr 29.
We developed and evaluated a rapid and simple multiplex microsphere assay for the quantification of specific IgG and IgA antibodies against meningococcal serogroup A, C, W, and Y capsular polysaccharides in serum and saliva. Meningococcal polysaccharides were conjugated to distinct magnetic carboxylated microspheres, and the performance of the assay was assessed using the CDC1992 standard meningococcal reference serum and a panel of serum and saliva samples. The standard curve was linear over an eight 3-fold dilution range in the IgG assay and a seven 3-fold dilution range in the IgA assay. No cross-reactivity was discovered, and the assay showed high specificity with ≥91% homologous inhibition and ≤11% heterologous inhibition for all serogroups and immunoglobulin classes. Lower limits of detections were ≤280 pg/ml for IgG and ≤920 pg/ml for IgA antibodies. The assay was reproducible, with a mean coefficient of variation of ≤5% for intra-assay duplicates, a mean coefficient of variation of ≤20% for interassay repeated analysis with different conjugations of microspheres, and a mean coefficient of variation within 25.8% for interoperator variation. The assay showed good correlation to the standard meningococcal polysaccharide enzyme-linked immunosorbent assay (ELISA) for detection of serum antibodies. This multiplex assay is robust and reliable and requires less sample volume, and less time and workload are needed than for ELISA, making this method highly relevant for serological and salivary investigations on the effect of meningococcal vaccines and for immunosurveillance studies.
我们开发并评估了一种快速简便的多重微球检测方法，用于定量血清和唾液中针对A、C、W和Y群脑膜炎球菌荚膜多糖的特异性IgG和IgA抗体。将脑膜炎球菌多糖与不同的磁性羧化微球偶联，并使用CDC1992标准脑膜炎球菌参考血清以及一组血清和唾液样本评估该检测方法的性能。在IgG检测中，标准曲线在8个3倍稀释范围内呈线性，在IgA检测中，标准曲线在7个3倍稀释范围内呈线性。未发现交叉反应，该检测方法对所有血清群和免疫球蛋白类别均显示出高特异性，同源抑制率≥91%，异源抑制率≤11%。IgG抗体的检测下限≤280 pg/ml，IgA抗体的检测下限≤920 pg/ml。该检测方法具有可重复性，批内重复检测的平均变异系数≤5%，不同微球偶联物的批间重复分析平均变异系数≤20%，不同操作人员之间的变异系数平均在25.8%以内。在检测血清抗体方面，该多重检测方法与标准脑膜炎球菌多糖酶联免疫吸附测定（ELISA）具有良好的相关性。这种多重检测方法稳健可靠，所需样本量较少，与ELISA相比所需时间和工作量更少，使得该方法在脑膜炎球菌疫苗效果的血清学和唾液学研究以及免疫监测研究中具有高度相关性。
