Bhide Ashlesha, Eldeeb Mohammed A, Pali Madhavi, Muthukumar Sriram, Prasad Shalini
Department of Bioengineering, The University of Texas at Dallas, Richardson, Texas 75080, United States.
EnLiSense LLC, 1813 Audubon Pond Way, Allen, Texas 75013, United States.
ACS Sens. 2023 Sep 22;8(9):3408-3416. doi: 10.1021/acssensors.3c00693. Epub 2023 Aug 29.
Exhaled breath condensate is an emerging source of inflammatory biomarkers suitable for the noninvasive detection of respiratory disorders. Current gold standard methods are highly invasive and pose challenges in sample collection during airway inflammation monitoring. Cytokine biomarkers are detectable in EBC at increased or decreased concentrations. IL-6, IL-1β, IL-8, and hs-CRP are characteristic biomarkers identified in respiratory disorders. We have demonstrated the promising outcomes of a 16-plexed electrochemical platform - READ 2.0 for the multiplexed detection of characteristic biomarkers in EBC. The sensor demonstrates dynamic ranges of 1-243 pg/mL with a lower detection limit of 1 pg/mL for IL-6 and IL-1β, while the detection range and limit of detection for IL-8 and hs-CRP is 1-150 pg/mL and 3 pg/mL, respectively. The detection accuracies for the biomarkers are in the range of ∼85 ± 15% to ∼100 ± 10%. The sensor shows a nonspecific response to similar cross-reacting biomarkers. Analytical validation of the sensor with ELISA as the standard reference generated a correlation of > 0.96 and mean biases of 10.9, 3.5, 17.4, and 3.9 pg/mL between the two methods for IL-6, IL-1β, IL-8, and hs-CRP, respectively. The precision of the sensor in detecting low biomarker concentrations yields a %CV of <7%. The variation in the sensor's response on repeat EBC sample measurements and within a 6 h duration is less than 10%. The READ 2.0 platform shows a promise that EBC-based biomarker detection can prove to be vital in predicting the severity and survival rates of respiratory disorders and serve as a reference point for monitoring EBC-based biomarkers.
呼出气冷凝物是一种新兴的炎症生物标志物来源,适用于呼吸系统疾病的无创检测。目前的金标准方法具有高度侵入性,并且在气道炎症监测期间的样本采集方面存在挑战。细胞因子生物标志物在呼出气冷凝物中可检测到浓度升高或降低。白细胞介素-6、白细胞介素-1β、白细胞介素-8和高敏C反应蛋白是在呼吸系统疾病中鉴定出的特征性生物标志物。我们已经证明了一种16通道电化学平台——READ 2.0在多重检测呼出气冷凝物中特征性生物标志物方面的良好结果。该传感器对白细胞介素-6和白细胞介素-1β的动态范围为1-243 pg/mL,检测下限为1 pg/mL,而白细胞介素-8和高敏C反应蛋白的检测范围和检测限分别为1-150 pg/mL和3 pg/mL。这些生物标志物的检测准确率在约85±15%至约100±10%的范围内。该传感器对类似的交叉反应生物标志物表现出非特异性反应。以酶联免疫吸附测定法作为标准参考对该传感器进行分析验证,两种方法之间白细胞介素-6、白细胞介素-1β、白细胞介素-8和高敏C反应蛋白的相关性>0.96,平均偏差分别为10.9、3.5、17.4和3.9 pg/mL。该传感器在检测低生物标志物浓度时的精密度产生的变异系数<7%。在重复测量呼出气冷凝物样本时以及在6小时内,该传感器响应的变化小于10%。READ 2.0平台表明,基于呼出气冷凝物的生物标志物检测在预测呼吸系统疾病的严重程度和生存率方面可能至关重要,并可作为监测基于呼出气冷凝物的生物标志物的参考点。