Multiplexed reverse-transcriptase quantitative polymerase chain reaction using plasmonic nanoparticles for point-of-care COVID-19 diagnosis
| Autor: | Nicole R. Blumenfeld, Michael Anne E. Bolene, Martin Jaspan, Abigail G. Ayers, Sabin Zarrandikoetxea, Juliet Freudman, Nikhil Shah, Angela M. Tolwani, Yuhang Hu, Terry L. Chern, James Rogot, Vira Behnam, Aditya Sekhar, Xinyi Liu, Bulent Onalir, Robert Kasumi, Abdoulaye Sanogo, Kelia Human, Kasey Murakami, Goutham S. Totapally, Mark Fasciano, Samuel K. Sia |
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| Rok vydání: | 2022 |
| Předmět: |
Reverse Transcriptase Polymerase Chain Reaction
SARS-CoV-2 Point-of-Care Systems Biomedical Engineering COVID-19 Bioengineering DNA-Directed RNA Polymerases Real-Time Polymerase Chain Reaction Condensed Matter Physics Sensitivity and Specificity Atomic and Molecular Physics and Optics COVID-19 Testing Humans Nanoparticles RNA Viral General Materials Science Electrical and Electronic Engineering Fluorescent Dyes |
| Zdroj: | Nature Nanotechnology. 17:984-992 |
| ISSN: | 1748-3395 1748-3387 |
| DOI: | 10.1038/s41565-022-01175-4 |
| Popis: | Quantitative polymerase chain reaction (qPCR) offers the capabilities of real-time monitoring of amplified products, fast detection, and quantitation of infectious units, but poses technical hurdles for point-of-care miniaturization compared with end-point polymerase chain reaction. Here we demonstrate plasmonic thermocycling, in which rapid heating of the solution is achieved via infrared excitation of nanoparticles, successfully performing reverse-transcriptase qPCR (RT-qPCR) in a reaction vessel containing polymerase chain reaction chemistry, fluorescent probes and plasmonic nanoparticles. The method could rapidly detect SARS-CoV-2 RNA from human saliva and nasal specimens with 100% sensitivity and 100% specificity, as well as two distinct SARS-CoV-2 variants. The use of small optical components for both thermocycling and multiplexed fluorescence monitoring renders the instrument amenable to point-of-care use. Overall, this study demonstrates that plasmonic nanoparticles with compact optics can be used to achieve real-time and multiplexed RT-qPCR on clinical specimens, towards the goal of rapid and accurate molecular clinical diagnostics in decentralized settings. |
| Databáze: | OpenAIRE |
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