School of Natural Sciences, University of Hull, Hull, UK.
Department of Materials and Environmental Chemistry, Stockholm University, Stockholm, Sweden.
Methods Mol Biol. 2024;2804:195-206. doi: 10.1007/978-1-0716-3850-7_12.
Clinical diagnostics of infectious diseases via nucleic acid amplification tests (NAATs) depend on a separate step of isolation of nucleic acids from cells/viruses embedded in complex biological matrices. The most recent example has been reverse transcription polymerase chain reaction (RT-PCR) for amplification and detection of SARS-CoV-2 RNA for COVID-19 diagnostics. Kits for RNA extraction and purification are commercially available; however, their integration with amplification systems is generally lacking, resulting in two separate steps, i.e., sample preparation and amplification. This makes NAATs more time-consuming, requiring skilled personnel, and can increase the likelihood of contamination. Here, we describe a setup and methodology to perform the quick extraction and detection of nucleic acids in an integrated manner. In particular, we focus on the use of an immiscible filtration device for capture, isolation, concentration, amplification, and colorimetric detection of SARS-CoV-2 RNA.
通过核酸扩增检测（NAATs）进行传染病的临床诊断依赖于从嵌入复杂生物基质中的细胞/病毒中分离核酸的单独步骤。最近的一个例子是用于 COVID-19 诊断的 SARS-CoV-2 RNA 扩增和检测的逆转录聚合酶链反应（RT-PCR）。用于 RNA 提取和纯化的试剂盒可商购获得；然而，它们通常与扩增系统不集成，导致两个单独的步骤，即样品制备和扩增。这使得 NAATs 更加耗时，需要熟练的人员，并且增加了污染的可能性。在这里，我们描述了一种设置和方法，以集成方式进行快速提取和检测核酸。特别是，我们专注于使用不混溶的过滤装置来捕获、分离、浓缩、扩增和比色检测 SARS-CoV-2 RNA。
