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dc.contributor.authorAlafeef, Maha
dc.contributor.authorMoitra, Parikshit
dc.contributor.authorDighe, Ketan
dc.contributor.authorPan, Dipanjan
dc.date.accessioned2021-05-28T17:01:20Z
dc.date.available2021-05-28T17:01:20Z
dc.date.issued2021-04-30
dc.identifier.urihttp://hdl.handle.net/10713/15844
dc.description.abstractThe global pandemic of coronavirus disease 2019 (COVID-19) highlights the shortcomings of the current testing paradigm for viral disease diagnostics. Here, we report a stepwise protocol for an RNA-extraction-free nano-amplified colorimetric test for rapid and naked-eye molecular diagnosis of COVID-19. The test employs a unique dual-prong approach that integrates nucleic acid (NA) amplification and plasmonic sensing for point-of-care detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), with a sample-to-assay response time of <1 h. The RNA-extraction-free nano-amplified colorimetric test utilizes plasmonic gold nanoparticles capped with antisense oligonucleotides (ASOs) as a colorimetric reporter to detect the amplified nucleic acid from the COVID-19 causative virus, SARS-CoV-2. The ASOs are specific for the SARS-CoV-2 N-gene, and binding of the ASOs to their target sequence results in the aggregation of the plasmonic gold nanoparticles. This highly specific agglomeration step leads to a change in the plasmonic response of the nanoparticles. Furthermore, when tested using clinical samples, the accuracy, sensitivity and specificity of the test were found to be >98.4%, >96.6% and 100%, respectively, with a detection limit of 10 copies/μL. The test can easily be adapted to diagnose other viral infections with a simple modification of the ASOs and primer sequences. It also provides a low-cost, rapid approach requiring minimal instrumentation that can be used as a screening tool for the diagnosis of COVID-19 at point-of-care settings in resource-poor situations. The colorimetric readout of the test can even be monitored using a handheld optical reader to obtain a quantitative response. Therefore, we anticipate that this protocol will be widely useful for the development of biosensors for the molecular diagnostics of COVID-19 and other infectious diseases. © 2021, The Author(s)en_US
dc.description.urihttps://doi.org/10.1038/s41596-021-00546-wen_US
dc.language.isoenen_US
dc.publisherSpringer Natureen_US
dc.relation.ispartofNature Protocolsen_US
dc.subjectRNA-extraction-free nano-amplified colorimetric testen_US
dc.subject.meshCOVID-19--diagnosisen_US
dc.subject.meshCOVID-19 Testing--methodsen_US
dc.subject.meshPoint-of-Care Testingen_US
dc.titleRNA-extraction-free nano-amplified colorimetric test for point-of-care clinical diagnosis of COVID-19en_US
dc.typeArticleen_US
dc.identifier.doi10.1038/s41596-021-00546-w
dc.identifier.pmid33931780
dc.source.countryEngland


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