Manually pressurized droplet digital PCR chip for rapid SARS-CoV-2 diagnostics

Author:

Elomaa Pinja12ORCID,Ojalehto Tuomas3ORCID,Kumar Darshan4ORCID,Jokinen Ville5ORCID,Saavalainen Päivi12ORCID

Affiliation:

1. Translational Immunology Research Program and Department of Medical and Clinical Genetics, University of Helsinki, Faculty of Medicine 1 , Haartmaninkatu 8, Helsinki 00290, Finland

2. Folkhälsan Research Center 2 , Helsinki, Finland

3. Aidian Oy 3 , Espoo, Finland

4. Aiforia Technologies Plc 4 , Helsinki, Finland

5. Department of Chemistry and Materials Science, Aalto University School of Chemical Engineering, Tietotie 3, 5 Espoo 02150, Finland

Abstract

Droplet digital PCR (ddPCR) is a technique in which PCR reaction is divided into thousands of nanoliter-sized droplets and has proven to be a great tool in virus diagnostics. Compared to the gold standard system quantitative real-time PCR (RT-qPCR), ddPCR functions particularly well when dealing with samples with low template counts, such as viral concentration. This feature makes the technique suitable for early detection of the virus. In this study, a novel portable PDMS ddPCR chip is introduced. The chip functions without external pumps using manual pressurization with a multichannel pipet. The created droplets are monodispersed and form a monolayer on the chip's collection chamber, from where they can be effortlessly imaged. Droplets were analyzed and counted using artificial intelligence. The use of the manually pressurized chip was demonstrated for a SARS-CoV-2 assay, which takes advantage of isothermal strand invasion-based amplification (SIBA) technology, allowing quick and accurate, even point-of-care analysis of the sample. The results demonstrate that SIBA assays can be divided into nanoliter-sized droplets and used as quantitative assays, giving an approximation of the samples' viral count.

Funder

Business Finland

Research Council of Finland

Orionin Tutkimussäätiö

Emil Aaltosen Säätiö

Publisher

AIP Publishing

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