Department of Biomedical Engineering, University of Houston, Houston, TX 77004, USA.
Biosensors (Basel). 2022 Sep 14;12(9):756. doi: 10.3390/bios12090756.
The inflammation biomarker Interleukin 6 (IL-6) exhibits a concentration of less than 7 pg/mL in healthy serum but increases 10-100-fold when inflammation occurs. Increased serum IL-6 has been reported in chronic diseases such as rheumatoid arthritis (RA), as well as in life-threatening acute illnesses such as sepsis and cytokine release syndrome (CRS). This work seeks to meet the demand for rapid detection of serum IL-6 both for rapid monitoring of chronic diseases and for triaging patients with acute illnesses. Following the optimization of several types of gold nanoparticles, membrane pore sizes, and buffer systems, an ultra-sensitive vertical flow assay (VFA) was engineered, allowing the detection of recombinant IL-6 in spiked buffer with a limit of detection (LoD) of 10 pg/mL and a reportable range of 10-10,000 pg/mL with a 15-min assay time. The detection of IL-6 in spiked pooled healthy serum exhibited an LoD of 3.2 pg/mL and a reportable range of 10-10,000 pg/mL. The VFA's stability was demonstrated over 1-day, two-week, four-week, and six-week storage durations at room temperature. The inter-operator CV and intra-operator CV were determined to be 14.3% and 15.2%, respectively. Three reference zones, high, low, and blank, were introduced into the cartridge to facilitate on-site semi-quantitative measurements across a 6-point semi-quantitative range. Finally, the performance of the IL-6 VFA was validated using 20 RA and 20 healthy control (HC) clinical serum samples, using ELISA as the gold standard platform. The ultra-sensitive, rapid IL-6 VFA could potentially be used to triage patients for intensive care, treatment adjustments, or for monitoring disease activity in inflammatory conditions.
炎症生物标志物白细胞介素 6（IL-6）在健康血清中的浓度低于 7pg/mL，但在发生炎症时会增加 10-100 倍。在类风湿关节炎（RA）等慢性疾病以及脓毒症和细胞因子释放综合征（CRS）等危及生命的急性疾病中，已报道血清 IL-6 增加。这项工作旨在满足对血清 IL-6 的快速检测的需求，既用于慢性疾病的快速监测，也用于对急性疾病患者进行分诊。在优化了几种类型的金纳米粒子、膜孔大小和缓冲系统之后，设计了一种超灵敏的垂直流动分析（VFA），允许以 10pg/mL 的检测限（LoD）和 10-10,000pg/mL 的报告范围检测缓冲液中重组 IL-6，测定时间为 15 分钟。在加标健康混合血清中检测 IL-6 的 LoD 为 3.2pg/mL，报告范围为 10-10,000pg/mL。VFA 在室温下的稳定性在 1 天、2 周、4 周和 6 周的储存期内得到了证明。确定了操作者间 CV 和操作者内 CV 分别为 14.3%和 15.2%。在试剂盒中引入了三个参考区，高、低和空白，以方便在 6 点半定量范围内进行现场半定量测量。最后，使用 ELISA 作为金标准平台，使用 20 个 RA 和 20 个健康对照（HC）临床血清样本验证了 IL-6 VFA 的性能。这种超灵敏、快速的 IL-6 VFA 可能被用于对患者进行重症监护、治疗调整或对炎症条件下的疾病活动进行监测。
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