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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: Apr 23, 2026

Two-Step Reverse Transcription Droplet Digital PCR Protocols for SARS-CoV-2 Detection and Quantification
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Bio-Rad and QIAGEN digital PCR platforms provide equivalent quantification for wastewater-based SARS-CoV-2

Thomas Clerkin1, Steph Smith1, Kevin Zhu2

  • 1Department of Earth, Marine, and Environmental Sciences, Institute of Marine Sciences, University of North Carolina at Chapel Hill, Morehead City, North Carolina, USA.

Applied and Environmental Microbiology
|April 22, 2026
PubMed
Summary

Digital PCR (dPCR) platforms from Bio-Rad and QIAGEN show equivalent performance for wastewater surveillance. This finding supports interchangeable use, aiding public health decision-making in pathogen monitoring.

Keywords:
CDC NWSSSARS-CoV-2digital PCRdroplet digital PCRmethod validationplatform comparisonpublic health surveillancequantitative detectionwastewater surveillancewastewater-based epidemiology

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Area of Science:

  • Environmental microbiology
  • Molecular diagnostics
  • Public health surveillance

Background:

  • Digital PCR (dPCR) offers advantages over quantitative PCR (qPCR) for wastewater surveillance, including precision and absolute quantification.
  • The Bio-Rad QX200 and QIAGEN QIAcuity are widely adopted dPCR platforms, but their comparative performance in wastewater applications is not well-established.

Purpose of the Study:

  • To conduct a blinded comparison of the Bio-Rad QX200 and QIAGEN QIAcuity dPCR platforms for wastewater surveillance.
  • To assess the quantitative equivalence and performance characteristics of both platforms across a range of SARS-CoV-2 concentrations.

Main Methods:

  • A blinded comparison of 93 archived wastewater influent samples was performed using Bio-Rad QX200 and QIAGEN QIAcuity platforms.
  • Samples were analyzed in triplicate for N1 and N2 SARS-CoV-2 gene targets and a bovine coronavirus control, stratified into low, medium, and high concentration bins.

Main Results:

  • Both dPCR platforms demonstrated statistically equivalent quantification (mean differences ≤0.12 log copies L⁻¹, R² > 0.93) across all targets and concentration levels.
  • Measurement variability was strongly correlated with wastewater treatment plant site (R² = 0.89), indicating sample matrix effects are more influential than the analytical platform.
  • Process limits of detection varied between platforms, with Bio-Rad QX200 being faster and QIAGEN QIAcuity having lower consumables cost.

Conclusions:

  • The Bio-Rad QX200 and QIAGEN QIAcuity platforms are quantitatively equivalent for wastewater surveillance, supporting their interchangeable use.
  • Platform selection should be based on laboratory-specific operational needs, such as throughput and cost.
  • These findings are critical for standardizing molecular methods in wastewater-based epidemiology and enabling data integration across diverse surveillance networks.