Influence of storage solution, temperature, assay time and concentration on RT-qPCR nucleic acid detection for SARS-CoV-2 detection of SARS-CoV-2 by the RT-qPCR

Biochem Biophys Res Commun. 2024 May 7:707:149726. doi: 10.1016/j.bbrc.2024.149726. Epub 2024 Mar 11.

Abstract

Real-time reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) is an important method for the early diagnosis of coronavirus disease 2019 (COVID-19). This study investigated the effects of storage solution, temperature and detection time on severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) nucleic acid detection by RT-qPCR. Various concentrations of SARS-CoV-2 were added to inactive and non-inactive storage solution and the viral suspensions were stored at various temperatures (room temperature, 4, -20 and -80 °C). Then, at five different detection time points, the Ct values were determined by RT-qPCR. Active and inactive storage solutions and storage temperature have a great impact on the detection of N gene of SARS-CoV-2 at different concentration corridors but have little impact on the ORF gene. The storage time has a greater impact on the N gene and ORF gene at high concentrations but has no effect on the two genes at low concentrations. In conclusion, storage temperature, storage time and storage status (inactivated, non-inactivated) have no effect on the nucleic acid detection of SARS-CoV-2 at the same concentration. For different concentrations of SARS-CoV-2, the detection of N gene is mainly affected.

Keywords: Detection time; RT-qPCR; SARS-CoV-2; Storage temperature; Viral storage solution.

MeSH terms

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

Substances

  • RNA, Viral