Comparison of Primer-Probe Sets among Different Master Mixes for Laboratory Screening of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2)

Biomed Res Int. 2020 Sep 25:2020:7610678. doi: 10.1155/2020/7610678. eCollection 2020.

Abstract

Background: There is a shortage of chemical reagents for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) diagnosis and a surge of SARS-CoV-2 cases, especially in limited-resource settings. Therefore, the combination of an optimal assay kit is necessary.

Methods: We compared the ability to screen SARS-CoV-2 among three primer-probe sets in two different master mixes, Invitrogen™ SuperScript™ III One-Step RT-PCR and LightCycler Multiplex RNA Virus Master.

Results: The assay with TIB-Molbiol, IDT, and Phu Sa sets for LightCycler Multiplex RNA Virus Master or Invitrogen™ SuperScript™ III One-Step RT-PCR showed positive results from a single reaction of triplicate in the three days of 4.8 copies per reaction. R squared and amplification efficiency were 0.97 and ranged from 107 to 108%, respectively.

Conclusions: Our findings indicated that TIB-Molbiol, IDT, and Phu Sa primer-probe sets could be beneficial for the laboratory screening of SARS-CoV-2 by RT-qPCR assay of E gene. There is a need to consider the combination of these reagent sets as a new strategy to increase the testing capacity of screening programs for COVID-19.

Publication types

  • Comparative Study

MeSH terms

  • Betacoronavirus / genetics*
  • Betacoronavirus / isolation & purification
  • COVID-19
  • COVID-19 Testing
  • COVID-19 Vaccines
  • Clinical Laboratory Techniques / methods*
  • Clinical Laboratory Techniques / statistics & numerical data
  • Coronavirus Infections / diagnosis*
  • Coronavirus Infections / epidemiology
  • Coronavirus Infections / virology
  • DNA Primers / genetics*
  • Humans
  • Multiplex Polymerase Chain Reaction / methods
  • Multiplex Polymerase Chain Reaction / statistics & numerical data
  • Pandemics
  • Pneumonia, Viral / diagnosis*
  • Pneumonia, Viral / epidemiology
  • Pneumonia, Viral / virology
  • RNA Probes / genetics*
  • RNA, Viral / genetics
  • Real-Time Polymerase Chain Reaction / methods
  • Real-Time Polymerase Chain Reaction / statistics & numerical data
  • Reverse Transcriptase Polymerase Chain Reaction / methods
  • Reverse Transcriptase Polymerase Chain Reaction / statistics & numerical data
  • SARS-CoV-2
  • Sensitivity and Specificity

Substances

  • COVID-19 Vaccines
  • Covid-19 aAPC vaccine
  • DNA Primers
  • RNA Probes
  • RNA, Viral