Oil immersed lossless total analysis system for integrated RNA extraction and detection of SARS-CoV-2

Nat Commun. 2021 Jul 14;12(1):4317. doi: 10.1038/s41467-021-24463-4.

Abstract

The COVID-19 pandemic exposed difficulties in scaling current quantitative PCR (qPCR)-based diagnostic methodologies for large-scale infectious disease testing. Bottlenecks include lengthy multi-step processes for nucleic acid extraction followed by qPCR readouts, which require costly instrumentation and infrastructure, as well as reagent and plastic consumable shortages stemming from supply chain constraints. Here we report an Oil Immersed Lossless Total Analysis System (OIL-TAS), which integrates RNA extraction and detection onto a single device that is simple, rapid, cost effective, and requires minimal supplies and infrastructure to perform. We validated the performance of OIL-TAS using contrived SARS-CoV-2 viral particle samples and clinical nasopharyngeal swab samples. OIL-TAS showed a 93% positive predictive agreement (n = 57) and 100% negative predictive agreement (n = 10) with clinical SARS-CoV-2 qPCR assays in testing clinical samples, highlighting its potential to be a faster, cheaper, and easier-to-deploy alternative for infectious disease testing.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • COVID-19 / diagnosis*
  • COVID-19 Nucleic Acid Testing / economics
  • COVID-19 Nucleic Acid Testing / instrumentation
  • COVID-19 Nucleic Acid Testing / methods*
  • Equipment Design
  • Humans
  • Molecular Diagnostic Techniques
  • Nasopharynx / virology
  • Nucleic Acid Amplification Techniques
  • RNA, Viral / genetics
  • RNA, Viral / isolation & purification
  • Reproducibility of Results
  • SARS-CoV-2 / genetics
  • SARS-CoV-2 / isolation & purification*
  • Sensitivity and Specificity
  • Time Factors
  • Virion / genetics
  • Virion / isolation & purification

Substances

  • RNA, Viral

Supplementary concepts

  • LAMP assay