Pickering Jerry W, Martins Thomas B, Greer Ryan W, Schroder M Carl, Astill Mark E, Litwin Christine M, Hildreth Stephen W, Hill Harry R
Associated Regional and University Pathologists Institute for Clinical and Experimental Pathology, Salt Lake City, UT 84108, USA.
Am J Clin Pathol. 2002 Apr;117(4):589-96. doi: 10.1309/lmch-c4q2-vfl9-3t1a.
We developed a multiplexed indirect immunofluorescent assay for antibodies to pneumococcal polysaccharides (PnPs) based on the Luminex multiple analyte profiling system (Luminex, Austin, TX). The assay simultaneously determines serum IgG concentrations to 14 PnPs serotypes: 1, 3, 4, 5, 6B, 7F, 8, 9N, 9V, 12F; 14, 18C, 19F, and 23F. To assess the specificity of the multiplexed assay for each individual serotype, inhibition-of-binding studies were conducted using adult serum samples obtained after pneumococcal vaccination. Except for the closely related serotypes 9V and 9N, we demonstrated inhibition by homologous serotypes of more than 95% and inhibition by heterologous serotypes of less than 15% for all 14 PnPs serotypes. There was, however, high heterologous inhibition of 50% or greater with some serotypes. These cross-reacting antibodies could not be removed by preabsorption with pneumococcal C-polysaccharide but were removed by additional preabsorption with serotype 22F polysaccharide. The multiplexed Luminex assay showed good overall agreement with a well-established enzyme-linked immunosorbent assay that is currently recommended for evaluation of pneumococcal vaccine immunogenicity.
我们基于Luminex多分析物检测系统（Luminex公司，得克萨斯州奥斯汀市）开发了一种用于检测肺炎球菌多糖（PnP）抗体的多重间接免疫荧光检测方法。该检测方法可同时测定血清中针对14种PnP血清型的IgG浓度，这些血清型分别为：1、3、4、5、6B、7F、8、9N、9V、12F、14、18C、19F和23F。为评估该多重检测方法对每种血清型的特异性，我们使用肺炎球菌疫苗接种后采集的成人血清样本进行了结合抑制研究。除了密切相关的血清型9V和9N外，我们发现所有14种PnP血清型的同源血清型抑制率均超过95%，而异源血清型抑制率均低于15%。然而，某些血清型存在50%或更高的异源抑制率。这些交叉反应抗体不能通过与肺炎球菌C多糖预吸收去除，但可通过额外与22F血清型多糖预吸收去除。多重Luminex检测方法与目前推荐用于评估肺炎球菌疫苗免疫原性的成熟酶联免疫吸附检测方法总体一致性良好。
