Tanak Ambalika Sanjeev, Jagannath Badrinath, Tamrakar Yashaswee, Muthukumar Sriram, Prasad Shalini
Department of Bioengineering, The University of Texas at Dallas, Richardson, TX, 75080, USA.
EnLiSense LLC, 1813 Audubon Pondway, Allen, TX, 75013, USA.
Anal Chim Acta X. 2019 Oct 3;3:100029. doi: 10.1016/j.acax.2019.100029. eCollection 2019 Nov.
In this work, we demonstrate a robust, dual marker, biosensing strategy for specific and sensitive electrochemical response of Procalcitonin and C-reactive protein in complex body fluids such as human serum and whole blood for the detection of sepsis. Enhanced sensitivity is achieved by leveraging the physicochemical properties of zinc oxide at the electrode-solution interface. Characterization techniques such as SEM, EDAX, AFM, FTIR and fluorescence microscopy were performed to ensure a suitable biosensing surface. The characteristic biomolecular interactions between the target analyte and specific capture probe is quantified through unique frequency signatures using non-faradaic electrochemical impedance spectroscopy (EIS). The developed biosensor demonstrated a detection limit of 0.10 ng mL for PCT in human serum and whole blood with an R of 0.99 and 0.98 respectively. CRP demonstrated a detection limit of 0.10 μg mL in human serum and whole blood with an R of 0.90 and 0.98 respectively. Cross-reactivity analysis demonstrated robust selectivity to PCT and CRP with negligible interaction to non-specific biomolecules. The novel aspect of this technology is the ability to fine-tune individual biomarkers response owing to the optimal frequency tuning capability. The developed biosensor requires an ultra-low sample volume of 10 μL without the need for sample dilution for rapid analysis. We envision the developed dual marker biosensor to be useful as a sepsis-screening device for prognostic monitoring.
在本研究中,我们展示了一种强大的双标记生物传感策略,用于在人血清和全血等复杂体液中对降钙素原(Procalcitonin)和C反应蛋白进行特异性和灵敏的电化学响应,以检测败血症。通过利用氧化锌在电极 - 溶液界面的物理化学性质实现了灵敏度的提高。采用扫描电子显微镜(SEM)、能量散射X射线光谱(EDAX)、原子力显微镜(AFM)、傅里叶变换红外光谱(FTIR)和荧光显微镜等表征技术来确保合适的生物传感表面。通过使用非法拉第电化学阻抗谱(EIS),通过独特的频率特征对目标分析物与特异性捕获探针之间的特征生物分子相互作用进行定量。所开发的生物传感器在人血清和全血中对PCT的检测限为0.10 ng/mL,相关系数(R)分别为0.99和0.98。CRP在人血清和全血中的检测限为0.10 μg/mL,相关系数分别为0.90和0.98。交叉反应分析表明,该传感器对PCT和CRP具有强大的选择性,与非特异性生物分子的相互作用可忽略不计。该技术的新颖之处在于,由于具有最佳的频率调谐能力,能够对各个生物标志物的响应进行微调。所开发的生物传感器仅需10 μL的超低样品体积,无需样品稀释即可进行快速分析。我们设想所开发的双标记生物传感器可作为一种用于预后监测的败血症筛查设备。