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Biochem Biophys Res Commun . Influence of storage solution, temperature, assay time and concentration on RT-qPCR nucleic acid detection for SARS-Co

tetano

tetano

Editor, Senior Moderator
  1. Biochem Biophys Res Commun
  4. . 2024 Mar 11:707:149726.
  5. doi: 10.1016/j.bbrc.2024.149726. Online ahead of print. 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
Lingxuan Su[SUP] 1 [/SUP], Yihan Lou[SUP] 1 [/SUP], Jiaxuan Li[SUP] 2 [/SUP], Haiyan Mao[SUP] 1 [/SUP], Jianhua Li[SUP] 1 [/SUP], Yi Sun[SUP] 1 [/SUP], Biaofeng Zhou[SUP] 1 [/SUP], Guangshang Wu[SUP] 2 [/SUP], Chen Huang[SUP] 3 [/SUP], Yanjun Zhang[SUP] 4 [/SUP], Keda Chen[SUP] 5 [/SUP]



Affiliations
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.

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