Department of Mechanical Engineering and Applied Mechanics, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA.
Analyst. 2021 May 21;146(10):3234-3241. doi: 10.1039/d0an02482a. Epub 2021 Apr 8.
Rapid and efficient biological sample preparation and pretreatment are crucial for highly sensitive, reliable and reproducible molecular detection of infectious diseases. Herein, we report a self-powered, integrated sample concentrator (SPISC) for rapid plasma separation, pathogen lysis, nucleic acid trapping and enrichment at the point of care. The proposed sample concentrator uses a combination of gravitational sedimentation of blood cells and capillary force for rapid, self-powered plasma separation. The pathogens (e.g., HIV virus) in separated plasma were directly lysed and pathogen nucleic acid was enriched by an integrated, flow-through FTA® membrane in the concentrator, enabling highly efficient nucleic acid preparation. The FTA® membrane of the SPISC is easy to store and transport at room temperature without need for uninterrupted cold chain, which is crucial for point of care sampling in resource-limited settings. The platform has been successfully applied to detect HIV virus in blood samples. Our experiments show that the sample concentrator can achieve a plasma separation efficiency as high as 95% and a detection sensitivity as low as 10 copies per 200 μL blood (∼100 copies per mL plasma) with variability less than 7%. The sample concentrator described is fully compatible with downstream nucleic acid detection and has great potential for early diagnostics, monitoring and management of infectious diseases at the point of care.
快速高效的生物样本制备和预处理对于传染病的高灵敏度、可靠和可重现的分子检测至关重要。在此,我们报告了一种自供电、集成的样品浓缩器(SPISC),用于在即时护理点快速分离血浆、病原体裂解、核酸捕获和浓缩。所提出的样品浓缩器结合使用血细胞的重力沉降和毛细作用力来实现快速、自供电的血浆分离。分离的血浆中的病原体(例如 HIV 病毒)直接在浓缩器中通过集成的、直通式 FTA®膜裂解,并且病原体核酸被浓缩,从而实现高效的核酸制备。SPISC 的 FTA®膜易于在室温下储存和运输,而不需要不间断的冷链,这对于资源有限环境下的即时护理采样至关重要。该平台已成功应用于检测血液样本中的 HIV 病毒。我们的实验表明,该样品浓缩器可以实现高达 95%的血浆分离效率和低至 10 拷贝/200μL 血液(约 100 拷贝/mL 血浆)的检测灵敏度,变异性小于 7%。所描述的样品浓缩器与下游核酸检测完全兼容,对于即时护理点传染病的早期诊断、监测和管理具有巨大的潜力。