Wastewater Monitoring with Digital PCR
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Testing Wastewater Samples for SARS-CoV-2 Using the Absolute Q
This study explores using the Absolute Q for screening of wastewater RNA samples with questionable or near limit qPCR results.
Fight Outbreaks with Early Warning
Viral RNA has been detected in environmental samples before symptoms appear.3 The Absolute Q facilitates accurate and sensitive detection of these rare molecules. The video below demonstrates reliable quantification of viral load without a standard curve for several sample types – including those collected from wastewater. In these studies, rare SARS-CoV-2 targets that were originally missed by qPCR were detected on the Absolute Q.
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- Sean C. Taylor et al. Droplet Digital PCR versus qPCR for gene expression analysis with low abundant targets: from variable nonsense to publication quality data. Scientific Reports. 2017 May 25;7(1):2409.
- Dong, Lianhua, et al. “Highly Accurate and Sensitive Diagnostic Detection of SARS-CoV-2 by Digital PCR.” MedRxiv, Cold Spring Harbor Laboratory Press, 1 Jan. 2020.
- Peccia, Jordan, et al. “SARS-CoV-2 RNA Concentrations in Primary Municipal Sewage Sludge as a Leading Indicator of COVID-19 Outbreak Dynamics.” MedRxiv, Cold Spring Harbor Laboratory Press, 1 Jan. 2020, doi.org/10.1101/2020.05.19.20105999.
1st demonstration of a true 1-step RT-PCR on a digital PCR platform in under 80 minutes
Low limit of detection for viral load quantification demonstrated down to 5 target copies per reaction among high human genomic DNA background
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.
- (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.
- 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