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高通量无试剂盒 RNA 提取流水线。

A high-throughput pipeline for scalable kit-free RNA extraction.

机构信息

Synthetic Biology for Clinical and Technological Innovation, National University of Singapore, 28 Medical Drive, Singapore, 117456, Singapore.

Synthetic Biology Translational Research Programme, Yong Loo Lin School of Medicine, National University of Singapore, 14 Medical Drive, Singapore, 117599, Singapore.

出版信息

Sci Rep. 2021 Dec 1;11(1):23260. doi: 10.1038/s41598-021-02742-w.

Abstract

An overreliance on commercial, kit-based RNA extraction in the molecular diagnoses of infectious disease presents a challenge in the event of supply chain disruptions and can potentially hinder testing capacity in times of need. In this study, we adapted a well-established, robust TRIzol-based RNA extraction protocol into a high-throughput format through miniaturization and automation. The workflow was validated by RT-qPCR assay for SARS-CoV-2 detection to illustrate its scalability without interference to downstream diagnostic sensitivity and accuracy. This semi-automated, kit-free approach offers a versatile alternative to prevailing integrated solid-phase RNA extraction proprietary systems, with the added advantage of improved cost-effectiveness for high volume acquisition of quality RNA whether for use in clinical diagnoses or for diverse molecular applications.

摘要

在传染病的分子诊断中过度依赖商业试剂盒的 RNA 提取,在供应链中断的情况下会带来挑战,并且在需要时可能会阻碍检测能力。在这项研究中,我们通过微型化和自动化将一种成熟的、强大的基于 TRIzol 的 RNA 提取方案改编为高通量格式。通过 RT-qPCR 检测 SARS-CoV-2 的检测对工作流程进行了验证,以说明其在不干扰下游诊断灵敏度和准确性的情况下的可扩展性。这种半自动、无试剂盒的方法为流行的集成固相 RNA 提取专利系统提供了一种多功能的替代方案,其优势在于无论用于临床诊断还是用于各种分子应用,都能以更高的性价比获得高质量的 RNA。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/39c5/8636496/0bfd282be2f7/41598_2021_2742_Fig1_HTML.jpg

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