tetano
Editor, Senior Moderator
ACS Nano
. 2021 Jun 9.
doi: 10.1021/acsnano.1c02494. Online ahead of print.
Extraction of Viral Nucleic Acids with Carbon Nanotubes Increases SARS-CoV-2 Quantitative Reverse Transcription Polymerase Chain Reaction Detection Sensitivity
Sanghwa Jeong[SUP] 1 2 [/SUP], Eduardo González-Grandío[SUP] 1 [/SUP], Nicole Navarro[SUP] 3 [/SUP], Rebecca L Pinals[SUP] 1 [/SUP], Francis Ledesma[SUP] 1 [/SUP], Darwin Yang[SUP] 1 [/SUP], Markita P Landry[SUP] 1 4 5 6 [/SUP]
Affiliations
Abstract
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 ∼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.
Keywords: DNA/RNA extraction; carbon nanotubes; nanosensors; nanotechnology; qPCR; viral diagnostics.
. 2021 Jun 9.
doi: 10.1021/acsnano.1c02494. Online ahead of print.
Extraction of Viral Nucleic Acids with Carbon Nanotubes Increases SARS-CoV-2 Quantitative Reverse Transcription Polymerase Chain Reaction Detection Sensitivity
Sanghwa Jeong[SUP] 1 2 [/SUP], Eduardo González-Grandío[SUP] 1 [/SUP], Nicole Navarro[SUP] 3 [/SUP], Rebecca L Pinals[SUP] 1 [/SUP], Francis Ledesma[SUP] 1 [/SUP], Darwin Yang[SUP] 1 [/SUP], Markita P Landry[SUP] 1 4 5 6 [/SUP]
Affiliations
- PMID: 34105936
- DOI: 10.1021/acsnano.1c02494
Abstract
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 ∼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.
Keywords: DNA/RNA extraction; carbon nanotubes; nanosensors; nanotechnology; qPCR; viral diagnostics.