Byrnes Samantha, Fan Andy, Trueb Jacob, Jareczek Francis, Mazzochette Mark, Sharon Andre, Sauer-Budge Alexis F, Klapperich Catherine M
Department of Biomedical Engineering, Boston University, Boston, MA 02215.
Anal Methods. 2013 Jul 7;5(13):3177-3184. doi: 10.1039/C3AY40162F.
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 µL), 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/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.
许多用于全球医学的新型且令人兴奋的便携式HIV病毒载量检测技术正在涌现。虽然向现场临床医生提供快速、等温且定量分子诊断信息的潜力很快将成为现实，但这些技术中的许多缺乏强大的前端样本净化和核酸制备功能。这样一种技术将能够实现许多不同的下游分子检测。在此，我们展示了一种便携式系统，该系统仅使用与在资源匮乏环境中储存和运输兼容的热稳定试剂，就能在室温下从小体积全血（70微升）中进行无离心机核酸提取。对模拟样本的定量逆转录聚合酶链反应（qRT-PCR）分析表明，检测下限为1000拷贝/毫升，能够检测四个数量级的病毒载量差异。该系统还可用于将提取的RNA储存在可拆卸的试剂盒上，在室温下保存长达一周，并且仅使用手动产生的气压即可操作。
