Sample-to-answer detection of miRNA from whole blood using thermally responsive alkane partitions

IF 10.5 1区生物学 Q1 BIOPHYSICS

Biosensors and Bioelectronics Pub Date : 2024-12-15 DOI:10.1016/j.bios.2024.117052

Evan H. Benke, David J. Boegner, Alejandra M. Bogusch, Ian M. White

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引用次数: 0

Abstract

Circulating miRNA offers a tremendous opportunity as a biomarker paradigm for many applications in disease diagnostics, including point-of-care diagnostics for global health needs. However, despite the numerous miRNA detection schemes reported, there still does not exist a solution for highly sensitive sample-to-answer detection of miRNA directly from complex samples, such as whole blood. We recently developed thermally responsive alkane partitions (TRAPs), which – when combined with magnetic microbeads – enable the complete assay automation from whole blood. Here we apply TRAPs with ligation-LAMP to automate the detection of miRNA in whole blood samples. MBs and a TRAP enable the automated purification of miRNA from blood, while a novel displacement-ligation method is utilized to trigger the ligation-LAMP reaction, which is streamlined into one step by a second TRAP. Using easily manufacturable TRAP-enabled assay cassettes and a custom low-cost handheld instrument, we report the specific detection of miR-155 at concentrations as low as 15 fM in whole blood with no intermediate steps by the user. This new approach creates the opportunity for point-of-care miRNA-based diagnostics with global health applications.

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利用热响应烷烃分区对全血中miRNA进行样本到应答检测。

循环 miRNA 作为一种生物标记范例，为疾病诊断的许多应用提供了巨大的机会，包括满足全球健康需求的护理点诊断。然而，尽管已报道了许多 miRNA 检测方案，但仍不存在直接从复杂样本（如全血）中进行高灵敏度 miRNA 样本到答案检测的解决方案。我们最近开发出了热响应烷烃隔板（TRAPs），它与磁性微珠相结合，可实现全血检测的完全自动化。在这里，我们将 TRAP 与连接-LAMP 结合使用，自动检测全血样本中的 miRNA。磁性微珠和 TRAP 可自动纯化血液中的 miRNA，同时利用新颖的置换-连接方法触发连接-LAMP 反应，通过第二个 TRAP 将该反应简化为一个步骤。利用易于制造的 TRAP 检测盒和定制的低成本手持仪器，我们报告了在全血中特异检测浓度低至 15 fM 的 miR-155，用户无需任何中间步骤。这种新方法为基于 miRNA 的全球健康应用点诊断创造了机会。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Biosensors and Bioelectronics 工程技术-电化学

CiteScore

20.80

自引率

7.10%

发文量

1006

审稿时长

29 days

期刊介绍： Biosensors & Bioelectronics, along with its open access companion journal Biosensors & Bioelectronics: X, is the leading international publication in the field of biosensors and bioelectronics. It covers research, design, development, and application of biosensors, which are analytical devices incorporating biological materials with physicochemical transducers. These devices, including sensors, DNA chips, electronic noses, and lab-on-a-chip, produce digital signals proportional to specific analytes. Examples include immunosensors and enzyme-based biosensors, applied in various fields such as medicine, environmental monitoring, and food industry. The journal also focuses on molecular and supramolecular structures for enhancing device performance.