Digital RT-PCR Chip method for detection of SARS-CoV-2 virus

J Immunol Methods. 2022 Oct:509:113339. doi: 10.1016/j.jim.2022.113339. Epub 2022 Aug 17.

Abstract

The "gold standard" method for detection of SARS-CoV-2 is the real time reverse transcription-polymerase chain reaction, but due to pre-analytical and technical limitations, biological samples with low viral load are not sometimes detected. For this purpose a digital RT-PCR method on-chip was developed for detection of the SARS-CoV-2 virus, using two TaqMan™ Assays for quantification of the N Protein (Nucleocapsid) and the S Protein (Spike), and the QuantStudio™ 3D Digital PCR instrument. The method was applied to assess the nasopharyngeal swabs of asymptomatic subjects recruited in the UNICORN Study. The digital RT-PCR method is characterized by a higher sensitivity than the RT-qPCR method, even if performed with the same TaqMan™, and could be a promising tool for SARS-CoV-2 viral load quantification.

Keywords: Chip; Digital RT-PCR; SARS-CoV-2; UNICORN; Viral load.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • COVID-19 Testing
  • COVID-19* / diagnosis
  • Humans
  • RNA, Viral / analysis
  • RNA, Viral / genetics
  • Real-Time Polymerase Chain Reaction / methods
  • Reverse Transcriptase Polymerase Chain Reaction
  • SARS-CoV-2* / genetics
  • Sensitivity and Specificity

Substances

  • RNA, Viral