Journal
ACS NANO
Volume 15, Issue 6, Pages 10309-10317Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acsnano.1c02494
Keywords
viral diagnostics; qPCR; DNA/RNA extraction; carbon nanotubes; nanotechnology; nanosensors
Categories
Funding
- IGI LGR ERA
- GlaxoSmithKline
- Burroughs Wellcome Fund Career Award at the Scientific Interface (CASI)
- Dreyfus foundation award
- Stanley Fahn PDF Junior Faculty Grant [PF-JFA-1760]
- Beckman Foundation Young Investigator Award
- NIH MIRA award
- NSF CAREER award
- NSF CBET award
- NSF CGEM award
- Sloan Foundation Award
- USDA BBT EAGER award
- USDA NIFA award
- Moore Foundation Award
- Cisco Research Center grant
- DARPA Young Investigator Award
- Office of Science, Office of Basic Energy Sciences, of the U.S. Department of Energy [DE-AC02-05CH11231]
- NSF [NSF DGE 1752814]
- Citris/Banatao Seed Funding
- FFAR Young Investigator award
- CZI investigator award
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The global SARS-CoV-2 pandemic has driven a surge in demand for rapid and efficient viral infection diagnostic tests, leading to a shortage in diagnostic test consumables. A new method using carbon nanotubes and ssDNA sequences has been developed to achieve high-yield extraction of viral nucleic acids, with a 100% extraction yield of target SARS-CoV-2 RNA. This method allows direct extraction of nucleic acids from human saliva, bypassing the need for further biofluid purification and commercial extraction kits.
The global SARS-CoV-2 coronavirus pandemic has led to a surging demand for rapid and efficient viral infection diagnostic tests, generating a supply shortage in diagnostic test consumables including nucleic acid extraction kits. Here, we develop a modular method for high-yield extraction of viral single-stranded nucleic acids by using capture ssDNA sequences attached to carbon nanotubes. Target SARS-CoV-2 viral RNA can be captured by ssDNA-nanotube constructs via hybridization and separated from the liquid phase in a single-tube system with minimal chemical reagents, for downstream quantitative reverse transcription polymerase chain reaction (RT-qPCR) detection. This nanotube-based extraction method enables 100% extraction yield of target SARS-CoV-2 RNA from phosphate-buffered saline in comparison to similar to 20% extraction yield when using a commercial silica-column kit. Notably, carbon nanotubes enable extraction of nucleic acids directly from 50% human saliva with a similar efficiency as achieved with commercial DNA/RNA extraction kits, thereby bypassing the need for further biofluid purification and avoiding the use of commercial extraction kits. Carbon nanotube-based extraction of viral nucleic acids facilitates high-yield and high-sensitivity identification of viral nucleic acids such as the SARS-CoV-2 viral genome with a reduced reliance on reagents affected by supply chain obstacles.
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