Development of an Extended-Specificity Multiplex Immunoassay for Detection of Streptococcus pneumoniae Serotype-Specific Antigen in Urine by Use of Human Monoclonal Antibodies
Development of an Extended-Specificity Multiplex Immunoassay for Detection of Streptococcus pneumoniae Serotype-Specific Antigen in Urine by Use of Human Monoclonal Antibodies
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DOI：
10.1128/cvi.00262-17
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发表时间：
2017-12-01
影响因子：
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通讯作者：
Fry, Norman K.
中科院分区：
文献类型：
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作者：
Eletu, Seyi D.;Sheppard, Carmen L.;Fry, Norman K.
目前的肺炎球菌疫苗涵盖目前描述的92种血清型中的10至23种最常见血清型。然而，随着肺炎球菌血清型疫苗使用的增加，血清型替代的风险和由非疫苗血清型引起的疾病的增加仍然存在。肺炎球菌感染的血清型监测在很大程度上依赖于培养技术，这种技术是不敏感的，特别是在非侵入性疾病的情况下。肺炎球菌特异性血清型尿液检测为侵袭性和非侵袭性疾病提供了另一种血清型方法。然而，先前描述的检测方法主要涵盖结合疫苗血清型，对流行的非疫苗血清型提供的信息很少，并且目前仅在一两个专业实验室中可用。我们的实验室开发了一种基于luminex的扩展范围抗原捕获测定法，用于检测尿液样本中的肺炎球菌血清型特异性抗原。该试验针对24种不同的血清型/血清群以及细胞壁多糖（CWP）和一些交叉反应血清型。我们报告说，该方法能够检测所有目标血清型和0.1 ng/ml的CWP，而某些血清型的检测浓度低至0.3 pg/ml。采用一组含非肺炎球菌细菌抗原的多糖阴性尿液标本，确定分析血清型特异性为98.4%。我们还报告了使用诊断为社区获得性肺炎或肺炎球菌病的患者的尿液标本组建立的临床敏感性为96.2%，特异性为89.9%。这种检测方法可以扩展到其他临床样本的检测，并且在许多从未获得培养物的肺炎球菌疾病病例中，具有极大改善血清型特异性监测的潜力。
Current pneumococcal vaccines cover the 10 to 23 most common serotypes of the 92 presently described. However, with the increased usage of pneumococcal-serotype-based vaccines, the risk of serotype replacement and an increase in disease caused by nonvaccine serotypes remains. Serotype surveillance of pneumococcal infections relies heavily on culture techniques, which are known to be insensitive, particularly in cases of noninvasive disease. Pneumococcal-serotypespecific urine assays offer an alternative method of serotyping for both invasive and noninvasive disease. However, the assays described previously cover mainly conjugate vaccine serotypes, give little information about circulating nonvaccine serotypes, and are currently available only in one or two specialist laboratories. Our laboratory has developed a Luminex-based extended -range antigen capture assay to detect pneumococcal-serotype-specific antigens in urine samples. The assay targets 24 distinct serotypes/serogroups plus the cell wall polysaccharide (CWP) and some cross -reactive serotypes. We report that the assay is capable of detecting all the targeted serotypes and the CWP at 0.1 ng/ml, while some serotypes are detected at concentrations as low as 0.3 pg/ml. The analytical serotype specificity was determined to be 98.4% using a panel of polysaccharide -negative urine specimens spiked with nonpneumococcal bacterial antigens. We also report clinical sensitivities of 96.2% and specificities of 89.9% established using a panel of urine specimens from patients diagnosed with community -acquired pneumonia or pneumococcal disease. This assay can be extended for testing other clinical samples and has the potential to greatly improve serotype-specific surveillance in the many cases of pneumococcal disease in which a culture is never obtained.