A simplified SARS-CoV-2 detection protocol for research laboratories

PLoS One. 2020 Dec 18;15(12):e0244271. doi: 10.1371/journal.pone.0244271. eCollection 2020.

Abstract

Widespread testing is required to limit the current public health crisis caused by the COVID-19 pandemic. Multiple tests protocols have been authorized by the food and drugs administration (FDA) under an emergency use authorization (EUA). The majority of these protocols are based on the gold-standard RT-qPCR test pioneered by the U.S. Centers for Disease Control and Prevention (CDC). However, there is still a widespread lack of testing in the US and many of the clinical diagnostics protocols require extensive human labor and materials that could face supply shortages and present biosafety concerns. Given the need to develop alternative reagents and approaches to provide nucleic-acid testing in the face of heightened demand and potential shortages, we have developed a simplified SARS-CoV-2 testing protocol adapted for its use in research laboratories with minimal molecular biology equipment and expertise. The protocol utilizes TRIzol to purify the viral RNA from different types of clinical specimens, requires minimal BSL-1 precautions and, given its high sensitivity, can be easily adapted to pooling samples strategies.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • COVID-19 / diagnosis*
  • COVID-19 Nucleic Acid Testing / methods*
  • COVID-19 Testing
  • Centers for Disease Control and Prevention, U.S.
  • HeLa Cells
  • Humans
  • Nasopharynx / virology
  • Nucleic Acid Amplification Techniques / methods*
  • Oropharynx / virology
  • Pandemics
  • RNA, Viral / isolation & purification*
  • Real-Time Polymerase Chain Reaction
  • SARS-CoV-2 / isolation & purification
  • Saliva / virology
  • Sensitivity and Specificity
  • United States

Substances

  • RNA, Viral

Grants and funding

This work was supported by the Florida Blue foundation grant “Developing Predictive Algorithms for COVID-19 Infection in FAU Health Care Workers”. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.