A portable, pressure driven, room temperature nucleic acid extraction and storage system for point of care molecular diagnostics
A portable, pressure driven, room temperature nucleic acid extraction and storage system for point of care molecular diagnostics
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DOI：
10.1039/c3ay40162f
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发表时间：
2013-01-01
影响因子：
3.1
通讯作者：
Klapperich, Catherine M.
中科院分区：
文献类型：
--
作者：
Byrnes, Samantha;Fan, Andy;Klapperich, Catherine M.
许多新的和令人兴奋的便携式HIV病毒载量检测技术正在出现，用于全球医学。虽然为该领域的临床医生提供快速、等温和定量分子诊断信息的可能性将很快成为现实，但这些技术中的许多技术缺乏用于样品清理和核酸制备的强大前端。这种技术将使许多不同的下游分子测定成为可能。在这里，我们提出了一种便携式系统，用于从少量全血（70 μ L）中进行无离心室温核酸提取，仅使用与低资源环境中的储存和运输兼容的热稳定试剂。模拟样本的定量逆转录酶聚合酶链反应（qRT-PCR）分析证明，检测下限为1000拷贝/mL，能够检测出四个数量级的病毒载量差异。该系统还可以用于在环境温度下将提取的RNA储存在可拆卸的盒上长达一周，并且可以仅使用手动产生的气压进行操作。
Many new and exciting portable HIV viral load testing technologies are emerging for use in global medicine. While the potential to provide fast, isothermal, and quantitative molecular diagnostic information to clinicians in the field will soon be a reality, many of these technologies lack a robust front end for sample clean up and nucleic acid preparation. Such a technology would enable many different downstream molecular assays. Here, we present a portable system for centrifuge-free room temperature nucleic acid extraction from small volumes of whole blood (70 mu L), using only thermally stable reagents compatible with storage and transport in low resource settings. Quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) analysis of simulated samples demonstrate a lower limit of detection of 1000 copies per mL, with the ability to detect differences in viral load across four orders of magnitude. The system can also be used to store extracted RNA on detachable cartridges for up to one week at ambient temperature, and can be operated using only hand generated air pressure.