Lathika S, Raj A, Sen A K
Fluid Systems Lab, Department of Mechanical Engineering, Indian Institute of Technology Madras Chennai India
Department of Mechanical Engineering, Indian Institute of Technology Patna Patna India.
RSC Adv. 2021 Oct 15;11(53):33770-33780. doi: 10.1039/d1ra05009e. eCollection 2021 Oct 8.
The development of a biosensor for rapid and quantitative detection of the dengue virus continues to remain a challenge. We report a lab-on-chip device that combines membrane-based blood plasma separation and a localized surface plasmon resonance (LSPR) based biosensor for on-chip detection of dengue NS1 antigen from a few drops of blood. The LSPR effect is realized by irradiating UV-NIR light having a spectral peak at 655 nm onto nanostructures fabricated thermal annealing of a thin metal film. We study the effect of the resulting metal nanostructures on the LSPR performance in terms of sensitivity and limit of detection, by annealing silver films at temperatures ranging from 100 to 500 °C. The effect of annealing temperature on the nanostructure size and uniformity and the resulting optical characteristics are investigated. Further, the binding between non-targeted blood plasma proteins and NS1-antibody-functionalized nanostructures on the LSPR performance is studied by considering different blocking mechanisms. Using a nanostructure annealed at 200 °C and 2X-phosphate buffer saline with 0.05% Tween-20 as the blocking buffer, from 10 μL of whole blood, the device can detect NS1 antigen at a concentration as low as 0.047 μg mL within 30 min. Finally, we demonstrate the detection of NS1 in the blood samples of dengue-infected patients and validate our results with those obtained from the gold-standard ELISA test.
开发一种用于快速定量检测登革热病毒的生物传感器仍然是一项挑战。我们报告了一种芯片实验室设备,该设备结合了基于膜的血浆分离技术和基于局域表面等离子体共振(LSPR)的生物传感器,用于从几滴血液中芯片检测登革热NS1抗原。LSPR效应是通过将光谱峰值在655nm的紫外-近红外光照射到通过对薄金属膜进行热退火制备的纳米结构上来实现的。我们通过在100至500°C的温度范围内对银膜进行退火,研究了所得金属纳米结构对LSPR性能在灵敏度和检测限方面的影响。研究了退火温度对纳米结构尺寸和均匀性以及由此产生的光学特性的影响。此外,通过考虑不同的封闭机制,研究了非靶向血浆蛋白与NS1抗体功能化纳米结构之间的结合对LSPR性能的影响。使用在200°C退火的纳米结构和含有0.05%吐温-20的2X磷酸盐缓冲盐水作为封闭缓冲液,从10μL全血中,该设备能够在30分钟内检测到低至0.047μg/mL浓度的NS1抗原。最后,我们展示了在登革热感染患者血液样本中NS1的检测,并将我们 的结果与通过金标准ELISA测试获得的结果进行了验证。