1. A high-throughput pipeline for scalable kit-free RNA extraction.
- Author
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Han P, Go MK, Chow JY, Xue B, Lim YP, Crone MA, Storch M, Freemont PS, and Yew WS
- Subjects
- COVID-19 virology, Humans, Molecular Diagnostic Techniques methods, RNA, Viral analysis, ROC Curve, COVID-19 diagnosis, COVID-19 Testing methods, High-Throughput Screening Assays methods, RNA, Viral genetics, RNA, Viral isolation & purification, Real-Time Polymerase Chain Reaction methods, SARS-CoV-2 genetics
- 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., (© 2021. The Author(s).)
- Published
- 2021
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