Absolute Quantitation of
SARS-CoV-2 with the Combinati Absolute Q dPCR Platform

Overview

Recently, RT-dPCR (reverse transcription digital PCR) has been shown to outperform traditional RT-qPCR in terms of sensitivity and accuracy in identifying patients who are infected with the SARS-CoV-2 virus. The Absolute Q digital PCR platform combines a streamlined workflow, less than 90 minutes of time-to-result and 4-color multiplexing to offer the best digital PCR solution for viral load quantification.

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Learn more about how the Absolute Q can accurately quantify as few as 5 viral targets per reaction using the 1-step |Q| SARS-CoV-2 Triplex Assay kit.

SARS-CoV-2 Reference Material Quantification

Reference materials for SARS-CoV-2 – both RNA and DNA – were quantified on the Absolute Q to demonstrate the platform’s performance. With high levels of consistency across instrument as well as high accuracy across multiple decades of concentrations, the Absolute Q digital PCR platform is the ideal system for reference material validation.

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1-Step RT-dPCR in under 90 minutes

In this technical note we showcase the simple workflow of the |Q| SARS-CoV-2 Triplex Assay Kit. With the Absolute Q’s integrated architecture, accurate quantitation of viral targets requires only 1 hands-on step and less than 90 minutes.

Sensitive detection of viral RNA targets

Reverse-transcription digital PCR (RT-dPCR) enables absolute quantification of viral targets without the need for a standard curve or use of reference materials. Here we demonstrate the Absolute Q’s sensitivity and consistency by robustly quantifying 5 copies of SARS-CoV-2 synthetic RNA molecules in a high background of human gDNA.

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Application Highlights

Digital PCR Turnaround Time

1st demonstration of a true 1-step RT-PCR on a digital PCR platform in under 80 minutes

Digital PCR Workflow

Low limit of detection for viral load quantification demonstrated down to 5 target copies per reaction among high human genomic DNA background

Digital PCR Multiplexing

Potential to enable more efficient quantification of viral targets with up to 4-color multiplexing

The COVID-19 pandemic has drawn heightened concern, with over eleven million positive SARS-CoV-2 cases confirmed worldwide by July 2020.¹ RT-qPCR currently serves as the clinical standard for the diagnosis of COVID-19. However in a recent study, it was demonstrated that digital PCR (dPCR) provided better sensitivity for identifying patients who ultimately were diagnosed with COVID-19.²

Reverse transcription digital PCR (RT-dPCR) is a valuable technique which enables improved consistency and lower limits of detection compared to qPCR. Quantification of extremely rare viral RNA target material is also possible without the need for a comparative standard curve.

Dynamic range of SARS-CoV-2 quantification on Absolute Q using |Q| SARS-CoV-2 Triplex Probe Assay

Results of dynamic range testing for SARS-CoV-2 RNA targets among 50 nanograms (~15,000 copies) of human genomic DNA. Individual points are plotted.

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Combinat’s Absolute Q platform is for research use only and has not been validated for use in diagnostic procedures.

Additional Resources

SARS-CoV-2 Reference Material Quantification on the Absolute Q dPCR Platform

SARS-CoV-2 Reference Material Quantification on the Absolute Q dPCR Platform

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1-Step RT-dPCR in under 80 Minutes on the Combinati Absolute Q

1-Step RT-dPCR in under 80 Minutes on the Combinati Absolute Q

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References

  1. (WHO), World Health Organization. “Coronavirus Disease (COVID-19) – Situation Report 169.” Coronavirus Disease (COVID-2019) Situation Reports, 7 July 2020, 10:00 CEST, www.who.int/docs/default-source/coronaviruse/situation-reports/20200707-covid-19-sitrep-169.pdf?sfvrsn=c6c69c88_2.
  2. Dong L, Zhou J, Niu C, et al. Highly accurate and sensitive diagnostic detection of SARS-CoV-2 by digital PCR. medRxiv. March 2020:2020.03.14.20036129. doi:10.1101/2020.03.14.20036129