Santopolo Giulia, Clemente Antonio, Aranda Maria, Socias Antonia, Del Castillo Alberto, Chica Aniceta, Borges Marcio, de la Rica Roberto
Multidisciplinary Sepsis Group, Health Research Institute of the Balearic Islands (IdISBa), 07120 Palma, Spain.
Chemistry Department, University of the Balearic Islands (UIB), 07122 Palma, Spain.
ACS Sens. 2021 Dec 24;6(12):4443-4450. doi: 10.1021/acssensors.1c01884. Epub 2021 Nov 18.
Hyperdegranulation of neutrophilic granulocytes is a common finding in sepsis that directly contributes to the heightened immune response leading to organ dysfunction. Currently, cell degranulation is detected by flow cytometry, which requires large infrastructure that is not always available at the point of care. Here, we propose a plasmonic assay for detecting the degranulation status of septic cells colorimetrically. It is based on triggering the aggregation of gold nanoparticles with cationic granule proteins. Cells from septic patients contain fewer granules and therefore release less cationic proteins than healthy cells. This results in red-colored assays than can be easily detected by eye. The assay can selectively detect cationic granule proteins even in the presence of an excess of unrelated proteins, which is key to detect degranulation with high specificity. Coupling this signal generation mechanism with a magnetic purification step enabled the identification of septic cells with the same performance as flow cytometry. This makes the proposed method a promising alternative for diagnosing sepsis in decentralized healthcare schemes.
中性粒细胞的过度脱颗粒是脓毒症中的常见现象,它直接导致免疫反应增强,进而引发器官功能障碍。目前,细胞脱颗粒是通过流式细胞术检测的,这需要大型设备,而这些设备在护理现场并不总是可用的。在此,我们提出一种等离子体检测法,用于比色检测脓毒症细胞的脱颗粒状态。它基于用阳离子颗粒蛋白触发金纳米颗粒的聚集。脓毒症患者的细胞含有的颗粒较少,因此释放的阳离子蛋白比健康细胞少。这导致比色检测呈红色,肉眼即可轻松检测到。该检测法即使在存在过量无关蛋白的情况下也能选择性地检测阳离子颗粒蛋白,这是高特异性检测脱颗粒的关键。将这种信号产生机制与磁纯化步骤相结合,能够以与流式细胞术相同的性能识别脓毒症细胞。这使得所提出的方法成为在分散式医疗保健方案中诊断脓毒症的一种有前景的替代方法。