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Related Concept Videos

Real Time RT-PCR02:57

Real Time RT-PCR

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Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...
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Related Experiment Video

Updated: Sep 18, 2025

Two-Step Reverse Transcription Droplet Digital PCR Protocols for SARS-CoV-2 Detection and Quantification
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Droplet Digital PCR or Real-Time PCR as a Method for Quantifying SARS-CoV-2 RNA in Plasma-Is There a Difference?

Beathe Kiland Granerud1,2,3, Mari Kaarbø2, Huda Al-Baldawi1

  • 1Institute of Clinical Medicine, University of Oslo, 0318 Oslo, Norway.

Viruses
|June 27, 2025
PubMed
Summary

This study compared reverse transcriptase real-time PCR (qRT-PCR) and digital droplet PCR (RT-ddPCR) for detecting SARS-CoV-2 RNA in COVID-19 patient plasma. Both methods showed comparable results for SARS-CoV-2 E-gene detection in plasma.

Keywords:
COVID-19E-geneRNAemiaRT-ddPCRSARS-CoV-2plasmaqRT-PCR

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Area of Science:

  • Molecular Biology
  • Virology
  • Infectious Diseases

Background:

  • Accurate detection of SARS-CoV-2 RNA in patient samples is crucial for COVID-19 diagnosis and management.
  • Quantitative reverse transcriptase real-time PCR (qRT-PCR) is a common method, but digital droplet PCR (RT-ddPCR) offers potential advantages in precision.
  • Comparing these methods for SARS-CoV-2 RNA quantification in clinical samples like plasma is essential for optimizing diagnostic strategies.

Purpose of the Study:

  • To evaluate and compare the effectiveness of qRT-PCR and RT-ddPCR for detecting SARS-CoV-2 RNA in blood plasma.
  • To assess the sensitivity and concordance between qRT-PCR and RT-ddPCR when quantifying the SARS-CoV-2 E-gene in COVID-19 patient plasma.

Main Methods:

  • Quantification of SARS-CoV-2 E-gene RNA was performed using both qRT-PCR and RT-ddPCR.
  • A total of 128 plasma samples from 70 hospitalized COVID-19 patients were analyzed.
  • Statistical analysis was employed to compare the sensitivity and concordance of the two molecular diagnostic methods.

Main Results:

  • Out of 128 plasma samples, 89 showed consistent SARS-CoV-2 RNA detection results between qRT-PCR and RT-ddPCR.
  • 39 samples exhibited discrepancies in viral RNA detection or quantification between the two methods.
  • RT-ddPCR often reported higher viral RNA quantities, but did not demonstrate a statistically significant superiority in sensitivity over qRT-PCR.

Conclusions:

  • Both qRT-PCR and RT-ddPCR provide comparable results for the detection of SARS-CoV-2 E-gene RNA in patient plasma.
  • While RT-ddPCR may yield higher viral load estimates, its sensitivity advantage over qRT-PCR was not definitively established in this study.
  • The findings suggest that both techniques are viable for SARS-CoV-2 RNA detection in plasma, with considerations for specific clinical applications.