肺炎链球菌
血清型
微生物学
抗体
多路复用
分析灵敏度
脑膜炎奈瑟菌
肺炎球菌多糖疫苗
多糖
肺炎球菌感染
异源的
生物
色谱法
分子生物学
化学
肺炎球菌病
细菌
医学
免疫学
抗生素
生物化学
生物信息学
遗传学
替代医学
病理
基因
作者
Gouri Lal,Paul Balmer,Elaine Stanford,Sarah A. Martin,R. E. Warrington,Ray Borrow
标识
DOI:10.1016/j.jim.2004.11.006
摘要
Serum IgG levels specific for Streptococcus pneumoniae are currently quantified using the ELISA. However, this method has significant limitations in that a separate test is required for each serotype-specific antibody. Here, we describe a rapid and simple method for the simultaneous quantitation of IgG to nine pneumococcal serotypes (1, 4, 5, 6B, 9V, 14, 18C, 19F, 23F). Pneumococcal polysaccharides were covalently attached to fluorescent microspheres and these beads were serologically assessed using 89-SF standard serum. The multiplex assay was linear over a 24-fold serum dilution range and comparison of monoplex and nonaplex assays revealed no evidence of bead interference. The nonaplex assay was shown to be specific with <30% heterologous inhibition occurring and assay sensitivity was high with the LOD ranging between 32.3 and 109.7 pg/ml. The assay was shown to have low inter- and intra-assay variability and conjugated microspheres were shown to remain stable over 12 months. When samples were assayed there was a good correlation of the nonaplex assay with the ELISA. Finally, four bead sets coated with meningococcal polysaccharides (serogroups A, C, Y and W135) were added into the nonaplex assay, with no effect on the pneumococcal or meningococcal IgG levels generated. The pneumococcal nonaplex assay will prove to be a valuable tool to aid in the evaluation of pneumococcal capsular polysaccharide-based vaccines.
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