Department of Chemistry and Biochemistry, Texas Tech University, Lubbock, Texas 79409, United States.
Clinical Research Institute, Texas Tech Health Sciences Center, Lubbock, Texas 79409, United States.
Anal Chem. 2023 Aug 29;95(34):12819-12825. doi: 10.1021/acs.analchem.3c01797. Epub 2023 Aug 9.
Sepsis is a serious medical condition that arises from a runaway response to an infection, which triggers the immune system to release chemicals into the bloodstream. This immune response can result in widespread inflammation throughout the body, which may cause harm to vital organs and, in more severe cases, lead to organ failure and death. Timely and accurate diagnosis of sepsis remains a challenge in analytical diagnostics. In this work, we have developed and validated a sepsis detection device, utilizing 3D printing technology, which incorporates multiple affinity separation zones. Our device requires minimal operator intervention and utilizes CD64, CD69, and CD25 as the biomarker targets for detecting sepsis in liquid biopsies. We assessed the effectiveness of our 3D-printed multizone cell separation device by testing it on clinical samples obtained from both septic patients ( = 35) and healthy volunteers ( = 8) and validated its performance accordingly. Unlike previous devices using poly(dimethyl siloxane), the 3D-printed device had reduced nonspecific binding for anti-CD25 capture, allowing this biomarker to be assayed for the first time in cell separations. Our results showed a statistically significant difference in cell capture between septic and healthy samples (with values of 0.0001 for CD64, CD69, and CD25), suggesting that 3D-printed multizone cell capture is a reliable method for distinguishing sepsis. A receiver operator characteristic (ROC) analysis was performed to determine the accuracy of the captured cell counts for each antigen in detecting sepsis. The ROC area under the curve (AUC) values for on-chip detection of CD64+, CD69+, and CD25+ leukocytes were 0.96, 0.92, and 0.88, respectively, indicating our diagnostic test matches clinical outcomes. When combined for sepsis diagnosis, the AUC value for CD64, CD69, and CD25 was 0.99, indicating an improved diagnostic performance due to the use of multiple biomarkers.
脓毒症是一种严重的医学病症，源于对感染的失控反应，这种反应会导致免疫系统将化学物质释放到血液中。这种免疫反应会导致全身广泛炎症，可能对重要器官造成伤害，在更严重的情况下，会导致器官衰竭和死亡。脓毒症的及时准确诊断仍然是分析诊断中的一个挑战。在这项工作中，我们开发并验证了一种利用 3D 打印技术的脓毒症检测设备，该设备包含多个亲和分离区。我们的设备需要最小的操作人员干预，并且利用 CD64、CD69 和 CD25 作为检测液体活检中脓毒症的生物标志物靶标。我们通过对来自脓毒症患者（n=35）和健康志愿者（n=8）的临床样本进行测试，评估了我们的 3D 打印多区细胞分离设备的有效性，并相应地验证了其性能。与以前使用聚二甲基硅氧烷的设备不同，3D 打印设备对 CD25 捕获的非特异性结合减少，从而首次能够在细胞分离中检测到该生物标志物。我们的结果表明，脓毒症和健康样本之间的细胞捕获存在统计学上的显著差异（CD64、CD69 和 CD25 的 值分别为 0.0001），这表明 3D 打印多区细胞捕获是区分脓毒症的一种可靠方法。进行了接收器操作特征（ROC）分析，以确定每个抗原对捕获细胞计数的准确性，以检测脓毒症。CD64+、CD69+和 CD25+白细胞在芯片上的检测的 ROC 曲线下面积（AUC）值分别为 0.96、0.92 和 0.88，表明我们的诊断测试与临床结果相符。当用于脓毒症诊断时，CD64、CD69 和 CD25 的 AUC 值为 0.99，表明由于使用了多个生物标志物，诊断性能得到了提高。
