Quantitative Real-Time RT-PCR Systems to Detect SARS-CoV-2

Methods Mol Biol. 2022:2511:89-97. doi: 10.1007/978-1-0716-2395-4_7.

Abstract

Since the outbreak of coronavirus disease 2019 (COVID-19) on the Diamond Princess cruise ship docked at Yokohama Port on February 3, 2020, real-time reverse transcription-polymerase chain reaction (RT-PCR) testing using nasopharyngeal swab samples from symptomatic and asymptomatic COVID-19 individuals has been the main way to detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA in almost all clinical laboratories in Japan. With the diffusion of sets containing the primers and probe, the gold standard real-time RT-PCR test has permeated throughout the country. To prevent the spread of infection, real-time RT-PCR testing is important to confirm whether people are positive, asymptomatic, or negative for COVID-19. Now, in addition to pharyngeal swab, saliva and blood samples can be used to detect SARS-CoV-2 RNA. Here, we introduce a clinical laboratory test performed using the High Pure viral nucleic acid kit and subsequent real-time RT-PCR system to detect SARS-CoV-2 RNA in serum, plasma, or whole blood in a hospital in Yokohama, Japan.

Keywords: Asymptomatic COVID-19 infection; Blood sample; Real-time RT-PCR; SARS-CoV-2 RNA.

MeSH terms

  • COVID-19* / diagnosis
  • Clinical Laboratory Techniques
  • Humans
  • RNA, Viral / genetics
  • Reverse Transcriptase Polymerase Chain Reaction
  • SARS-CoV-2* / genetics

Substances

  • RNA, Viral