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Related Concept Videos

Real Time RT-PCR02:57

Real Time RT-PCR

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: May 16, 2026

Quantification of Adeno-Associated Viral Genomes in Purified Vector Samples by Digital Droplet Polymerase Chain Reaction
04:43

Quantification of Adeno-Associated Viral Genomes in Purified Vector Samples by Digital Droplet Polymerase Chain Reaction

Published on: October 11, 2024

Digital PCR for Absolute Quantification of Adenoviruses.

Mikayla Quinton1, Omar Abdullah1, Amary Fall1

  • 1Division of Medical Microbiology, Department of Pathology, Johns Hopkins School of Medicine, Baltimore, Maryland.

The Journal of Molecular Diagnostics : JMD
|May 14, 2026
PubMed
Summary

Digital PCR (dPCR) offers accurate and reproducible quantification of human adenovirus (HAdV) viral loads. This study validates the Qiagen QIAcuityDx dPCR platform, showing its utility for standardizing molecular testing.

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Published on: July 14, 2023

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Last Updated: May 16, 2026

Quantification of Adeno-Associated Viral Genomes in Purified Vector Samples by Digital Droplet Polymerase Chain Reaction
04:43

Quantification of Adeno-Associated Viral Genomes in Purified Vector Samples by Digital Droplet Polymerase Chain Reaction

Published on: October 11, 2024

A Validatable Droplet Digital Polymerase Chain Reaction Assay for the Detection of Adeno-Associated Viral Vectors in Bioshedding Studies of Tears
07:43

A Validatable Droplet Digital Polymerase Chain Reaction Assay for the Detection of Adeno-Associated Viral Vectors in Bioshedding Studies of Tears

Published on: July 14, 2023

Area of Science:

  • Molecular Biology
  • Virology
  • Clinical Diagnostics

Background:

  • Quantitative real-time PCR (qPCR) assays exhibit significant variability in viral load reporting due to inconsistent calibration materials, assay designs, and extraction methods.
  • Lack of standardization in qPCR assays complicates accurate quantification and comparison of viral loads, particularly for adenoviruses (HAdV).
  • Digital PCR (dPCR) offers a potential solution for absolute quantification and improved standardization in molecular diagnostics.

Purpose of the Study:

  • To evaluate the Qiagen QIAcuityDx digital PCR (dPCR) platform for the absolute quantification of adenoviruses.
  • To compare the performance of the Qiagen QIAcuityDx dPCR platform against Bio-Rad droplet dPCR (ddPCR) and standard-of-care quantitative real-time PCR (qPCR) assays.
  • To assess the impact of adenovirus genotype on quantification precision across different platforms.

Main Methods:

  • Evaluation of the Qiagen QIAcuityDx dPCR platform using commercially available quality control materials, calibrators, and clinical samples.
  • Comparative analysis with Bio-Rad droplet dPCR (ddPCR) and standard-of-care qPCR assays.
  • Testing of human adenovirus (HAdV) verification panels and control materials (Bio-Rad HAdV E4, ZeptoMetrix HAdV B3) on both dPCR systems.
  • Analysis of remnant clinical samples representing sixteen HAdV genotypes in serial dilutions across all tested platforms.

Main Results:

  • Both dPCR systems demonstrated excellent correlation and equivalent analytical sensitivity at 1000 copies/mL.
  • Strong agreement was observed across all assays for viral loads ≥1000 copies/mL, irrespective of HAdV genotype.
  • The Qiagen QIAcuityDx platform showed a minor average bias of -0.31 log copies/mL (SD 0.44) relative to qPCR.
  • Both dPCR systems exhibited high concordance with each other and with standard-of-care quantification, despite minor platform differences.

Conclusions:

  • Digital PCR (dPCR) enables accurate and reproducible quantification of HAdV viral loads.
  • The Qiagen QIAcuityDx dPCR platform is suitable for absolute quantification and supports its utility in calibration and standardization for clinical molecular testing.
  • dPCR platforms provide a standardized approach for HAdV quantification, overcoming limitations of traditional qPCR methods.