Kofoed Kristian, Schneider Uffe Vest, Scheel Troels, Andersen Ove, Eugen-Olsen Jesper
Clinical Research Unit, Copenhagen University Hospital Hvidovre, Hvidovre, Denmark.
Clin Chem. 2006 Jul;52(7):1284-93. doi: 10.1373/clinchem.2006.067595. Epub 2006 May 11.
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.
We used a combination of in-house and commercially available multiplex immunoassays based on Luminex xMAP technology to assay biomarkers of potential interest in EDTA-plasma samples.
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-1beta (IL-1beta), IL-6, IL-8, granulocyte/macrophage colony-stimulating factor, and tumor necrosis factor-alpha 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.
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.
脓毒症是一种常见且往往致命的疾病。由于脓毒症可由多种不同生物体引起，因此非常需要有助于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能会提供额外信息以补充目前使用的标志物。
我们使用基于Luminex xMAP技术的内部和市售多重免疫测定组合，来检测乙二胺四乙酸（EDTA）血浆样本中潜在感兴趣的生物标志物。
开发并在内部验证了一种用于检测可溶性尿激酶型纤溶酶原激活物受体（suPAR）、髓系细胞触发受体-1（sTREM-1）和巨噬细胞移动抑制因子（MIF）的三联检测法。这种三联检测法被添加到市售的白细胞介素-1β（IL-1β）、IL-6、IL-8、粒细胞/巨噬细胞集落刺激因子和肿瘤坏死因子-α人类细胞因子检测组中。组合检测时未观察到交叉反应。八联检测法、五细胞因子检测组、三种内部单联检测法以及suPAR酶联免疫吸附测定（ELISA）所获得的值之间的相关性在0.86至0.99之间。批内和批间平均变异系数（CV）分别为8.0%和11%。suPAR、sTREM-1和MIF校准品的回收率分别为108%、88%和51%。在通过血培养确诊的10例细菌性脓毒症患者采集的血浆中，与健康对照相比，该检测法检测到所有8种分析物的浓度均显著升高。
市售的xMAP检测组可用感兴趣的标志物进行扩展。联合多重检测法能够以高重现性测量这8种分析物。xMAP技术是一种用于联合检测传统细胞因子和新标志物的有吸引力的工具。
