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Method for Simultaneous Measurement of Antibodies to 23 Pneumococcal Capsular Polysaccharides

机译:同时测量23种肺炎球菌荚膜多糖抗体的方法

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We describe a fluorescent covalent microsphere immunoassay (FCMIA) method for the simultaneous (multiplexed) measurement of immunoglobulin G (IgG) antibodies to 23 pneumococcal capsular polysaccharide (PnPS) serotypes present in the pneumococcal polysaccharide vaccine (PPV23) licensed by the Food and Drug Administration, i.e., PnPSs 1, 2, 3, 4, 5, 6B, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19A, 19F, 20, 22F, 23F, and 33F. In addition, the assay incorporates an internal control that allows for contemporaneous evaluation of the effectiveness of pneumococcal cell wall polysaccharide (C-PS) preadsorption and a second control of PnPS 25 (which is not present in any polysaccharide or conjugate vaccine), which can be used to evaluate interassay reproducibility (useful for pre- versus postvaccination studies). The FCMIA was standardized with U.S. reference antipneumococcal serotype standard serum 89S-2. Preadsorption of 89S-2 with each PnPS and C-PS yielded homologous inhibition for serotypes 1, 6B, 9N, 9V, 11A, 12F,14, 15B, 18C, 19A, 19F, 20, 22F, 25, and 33F; heterologous inhibition for serotypes 9V, 10A, 11A, 12F, 15B, 17F, 20, and 23F; and neither homologous nor heterologous inhibition for serotypes 2, 3, 4, and 5. The minimum detectable concentrations for the 24 multiplexed (PnPS and C-PS) FCMIAs ranged from 20 pg/ml for PnPS 3 to 600 pg/ml for PnPS 14. The PnPS FCMIA method has numerous benefits over enzyme-linked immunosorbent assays commonly used to measure anti-PnPS-specific IgG levels, including increased speed, smaller sample volumes, equivalent or better sensitivity, and increased dynamic range.
机译:我们描述了一种荧光共价微球免疫测定(FCMIA)方法,用于同时(多重)测量针对23种肺炎球菌荚膜多糖(PnPS)血清型的免疫球蛋白G(IgG)抗体,该抗体存在于食品和药物管理局许可的肺炎球菌多糖疫苗(PPV23)中即PnPS 1、2、3、4、5、6B,7F,8、9N,9V,10A,11A,12F,14、15B,17F,18C,19A,19F,20、22F,23F和33F 。此外,该测定法还包含一个内部对照,可以同时评估肺炎球菌细胞壁多糖(C-PS)预吸附的有效性和另一个PnPS 25对照(在任何多糖或结合疫苗中都不存在),它可以可用于评估测定间的可重复性(用于疫苗接种前和疫苗接种后研究)。 FCMIA用美国参考抗肺炎球菌血清型标准血清89S-2标准化。每种PnPS和C-PS对89S-2的前吸附产生对血清型1、6B,9N,9V,11A,12F,14、15B,18C,19A,19F,20、22F,25和33F的同源抑制;对血清型9V,10A,11A,12F,15B,17F,20和23F的异源抑制;且对血清型2、3、4和5均无同源或异源抑制。24种多重(PnPS和C-PS)FCMIA的最低可检测浓度范围从PnPS 3的20 pg / ml到PnPS 14的600 pg / ml。与通常用于测量抗PnPS特异性IgG水平的酶联免疫吸附测定相比,PnPS FCMIA方法具有许多优势,包括提高速度,减少样品量,等效或更好的灵敏度以及增加的动态范围。

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