Development and validation of a multiplex add-on assay for sepsis biomarkers using xMAP technology
Development and validation of a multiplex add-on assay for sepsis biomarkers using xMAP technology
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DOI：
10.1373/clinchem.2006.067595
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发表时间：
2006-07-01
影响因子：
9.3
通讯作者：
Eugen-Olsen, Jesper
中科院分区：
文献类型：
--
作者：
Kofoed, Kristian;Schneider, Uffe Vest;Eugen-Olsen, Jesper
背景：脓毒症是一种。一种常见的致命疾病。由于脓毒症可以由许多不同的生物体引起，因此高度保证可以帮助诊断脓毒症和监测治疗效果的生物标志物。新的脓毒症标志物可以提供额外的信息，以补充目前使用的markers.Methods：我们使用了一个组合的内部和商业上可用的多重免疫分析的基础上Luminex（R）xMAP技术，以测定生物标志物的潜在利益的EDTA-血浆样本。可溶性尿激酶纤溶酶原激活物受体（suPAR）、髓样细胞上表达的可溶性触发受体-1（斯特雷姆-1）巨噬细胞迁移抑制因子（MIF）是在内部开发和验证的。将该3重测定加入到市售白细胞介素-1 β（IL-1 β）、IL-6、IL-8、粒细胞/巨噬细胞集落刺激因子和肿瘤坏死因子-a人细胞因子组中。当组合检测时，未观察到交叉反应性。8-plex、5-细胞因子组、3种内部1-plex测定和suPAR ELISA获得的值之间的相关性范围为0.86至0.99。平均运行内和运行间CV分别为8.0%和11%。suPAR、斯特雷姆-1和MIF校准品的回收率分别为108%、88%和51%。在血浆中收集的10例细菌性脓毒症患者的血培养证实，检测显着增加的浓度与健康controls.Conclusions相比，所有8种分析物：市售的xMAP面板可以扩展与标记的兴趣。组合的多重检测可以高重现性地测量8种分析物。xMAP技术是一种有吸引力的工具，用于分析传统细胞因子与新标记物的组合。(c)2006年美国临床化学协会。
Background: Sepsis is a. common and often fatal disease. Because sepsis can be caused by many different organisms, biomarkers that can aid in diagnosing sepsis and monitoring treatment efficacy are highly warranted. New sepsis markers may provide additional information to complement the currently used markers.Methods: We used a combination of in-house and commercially available multiplex immunoassays based on Luminex (R) xMAP technology to assay biomarkers of potential interest in EDTA-plasma samples.Results: A 3-plex assay for soluble urokinase plasminogen activator receptor (suPAR), soluble triggering receptor expressed on myeloid cells-1 (sTREM-1), and macrophage migration inhibiting factor (MIF) was developed and validated in-house. This 3-plex assay was added to a commercially available interleukin-1 beta (IL-1 beta), IL-6, IL-8, granulocyte/macrophage colony-stimulating factor, and tumor necrosis factor-a human cytokine panel. No cross-reactivity was observed when the assays were combined. Correlation between values obtained with the 8-plex, the 5-cytokine panel, the 3 in-house 1-plex assays, and a suPAR ELISA ranged from 0.86 to 0.99. Mean within- and between-run CVs were 8.0% and 11%, respectively. Recoveries of suPAR, sTREM-1, and MIF calibrators were 108%, 88%, and 51%, respectively. In plasma collected from 10 patients with bacterial sepsis confirmed by blood culture, the assay detected significantly increased concentrations of all 8 analytes compared with healthy controls.Conclusions: A commercially available xMAP panel can be expanded with markers of interest. The combined multiplex assay can measure the 8 analytes with high reproducibility. The xMAP technology is an appealing tool for assaying conventional cytokines in combination with new markers. (c) 2006 American Association for Clinical Chemistry.