Hairpin-Spherical Nucleic Acids for Diagnosing COVID-19: a Simple Method to Generalize the Conventional PCR for Molecular Assays
| Autor: | Masoumeh Hasani, Razieh Ezati, Farid Azizi Jalilian, Abbas Karami |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
DNA polymerase
Metal Nanoparticles 010402 general chemistry Polymerase Chain Reaction Sensitivity and Specificity 01 natural sciences Article Analytical Chemistry law.invention chemistry.chemical_compound Recognition sequence law Nucleic Acids TaqMan Humans Pandemics Polymerase chain reaction biology SARS-CoV-2 010401 analytical chemistry COVID-19 RNA Molecular biology 0104 chemical sciences Reverse transcription polymerase chain reaction chemistry Nucleic acid biology.protein RNA Viral Gold DNA |
| Zdroj: | Analytical Chemistry |
| ISSN: | 1520-6882 0003-2700 |
| DOI: | 10.1021/acs.analchem.1c01515 |
| Popis: | The COVID-19 pandemic revealed during the first global wave of this infectious disease that mass diagnostic testing was necessary to more rapidly detect infection in patients and control the pandemic. Therefore, extra research efforts to develop reliable and more accessible techniques for disease diagnosis are of supreme importance. Here, a target-responsive assembly of gold nanoparticle-core hairpin-spherical nucleic acids (AuNP-core H-SNAs) was implemented to modify the conventional polymerase chain reaction (PCR) assay for the "naked-eye" colorimetric detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA. Two hairpin DNA ligands are designed based on the two highly conserved regions within N and E genes of SARS-CoV-2 RNA by positioning two short palindromic arms (stem) on either side of a recognition sequence (loop). In the presence of a sequence-specific probe (activator), hairpin DNAs anchored to the surface of AuNPs unfold and expose the palindromic ends to the DNA-directed assembly of AuNPs. The sequence of the activator probes was chosen to be identical to the TaqMan probe in a real-time reverse transcription PCR (RT-PCR) assay for specifically targeting the N and E genes of SARS-CoV-2 RNA. They may either be degraded by the 5'-exonuclease activity of DNA polymerase during PCR cycles or stay intact depending on the presence or absence of the target template in the sample, respectively. Post-addition of H-SNA solutions to the final PCR products of some preconfirmed clinical samples for COVID-19 generated naked-eye-observable red and blue colors for positive and negative cases, respectively, with similar sensitivity to that of the real-time RT-PCR method. |
| Databáze: | OpenAIRE |
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