"Sample pooling of RNA extracts to speed up SARS-CoV-2 diagnosis using CDC FDA EUA RT-qPCR kit"

Virus Res. 2020 Dec:290:198173. doi: 10.1016/j.virusres.2020.198173. Epub 2020 Sep 24.

Abstract

Background: The CDC protocol for SARS-CoV2 RT-PCR diagnosis (2019-nCoV CDC kit) is considered a gold standard worldwide; based on three different FAM probes (N1 and N2 for viral detection; RP for RNA extraction quality control), three reactions per sample are needed for SARS-CoV-2 diagnosis.

Results: We herein describe a sample pooling protocol: pooling 3 RNA extractions into a single PCR reaction; we tested this protocol with 114 specimens grouped in 38 pools and found no significant differences for N1 and N2 Ct values between pool and single sample PCR reaction.

Conclusion: This pool of three protocol has a sensitivity of 100 % compared to the standard single sample protocol. For a typical 96-well plate, this pool assay allows 96 samples processing, speeding up diagnosis and reducing cost while maintaining clinical performance, particularly useful for SARS-CoV-2 diagnosis at developing countries.

Keywords: CDC; Pools; RT-qPCR; SARS-CoV-2.

Publication types

  • Evaluation Study

MeSH terms

  • COVID-19 / diagnosis*
  • COVID-19 Nucleic Acid Testing* / economics
  • Diagnostic Tests, Routine
  • Humans
  • Nasopharynx / virology
  • RNA, Viral / genetics
  • Reagent Kits, Diagnostic
  • Real-Time Polymerase Chain Reaction
  • SARS-CoV-2 / genetics
  • SARS-CoV-2 / isolation & purification*
  • Sensitivity and Specificity
  • Specimen Handling / methods*
  • Specimen Handling / standards
  • Time Factors

Substances

  • RNA, Viral
  • Reagent Kits, Diagnostic