Jongwon Lim, Joanne Hwang, Hyegi Min, Matthew Wester, Chansong Kim, Enrique Valera, Hyun Joon Kong, Rashid Bashir
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引用次数: 0
Abstract
The gold standard for diagnosing viruses such as the Hepatitis B Virus has remained largely unchanged, relying on conventional methods involving extraction, purification, and polymerase chain reaction (PCR). This approach is hindered by limited availability, as it is time-consuming and requires highly trained personnel. Moreover, it suffers from low recovery rates of the nucleic acid molecules for samples with low copy numbers. To address the challenges of complex instrumentation and low recovery rate of DNA, a drying process coupled with thermal treatment of whole blood is employed, resulting in the creation of a dried blood matrix characterized by a porous structure with a high surface-to-volume ratio where it also inactivates the amplification inhibitors present in whole blood. Drawing on insights from Brunauer-Emmett-Teller (BET)- Barrett-Joyner-Halenda (BJH) analysis, scanning electron microscopy (SEM), and fluorescence recovery after photobleaching (FRAP), detection assay is devised for HBV, as a demonstration, from whole blood with high recovery of DNA and simplified instrumentation achieving a limit of detection (LOD) of 10 IU mL-1. This assay can be completed in <1.5 h using a simple heater, can be applied to other DNA viruses, and is expected to be suitable for point-of-care, especially in low-resource settings.
诊断乙型肝炎病毒等病毒的黄金标准在很大程度上一直未变,仍然依赖于提取、纯化和聚合酶链反应(PCR)等传统方法。这种方法由于耗时长,需要训练有素的人员,因此可用性有限。此外,对于拷贝数较低的样本,核酸分子的回收率也很低。为了解决仪器复杂和 DNA 回收率低的难题,我们采用了一种干燥工艺,并对全血进行热处理,从而产生一种干燥的血液基质,这种基质具有多孔结构,表面积与体积比很高,还能使全血中的扩增抑制剂失活。借鉴布鲁瑙尔-艾美特-泰勒(BET)-巴雷特-乔伊纳-哈伦达(BJH)分析、扫描电子显微镜(SEM)和光漂白后荧光恢复(FRAP)的见解,设计出了检测 HBV 的分析方法,作为示范,从全血中检测 DNA 的回收率很高,仪器简化,检测限(LOD)为 10 IU mL-1。该检测可在以下时间内完成
期刊介绍:
Advanced Healthcare Materials, a distinguished member of the esteemed Advanced portfolio, has been dedicated to disseminating cutting-edge research on materials, devices, and technologies for enhancing human well-being for over ten years. As a comprehensive journal, it encompasses a wide range of disciplines such as biomaterials, biointerfaces, nanomedicine and nanotechnology, tissue engineering, and regenerative medicine.