Conventional PCR Assisted Single-Component Assembly of Spherical Nucleic Acids for Simple Colorimetric Detection of SARS-CoV-2
| Autor: | Masoumeh Hasani, Razieh Ezati, Farid Azizi Jalilian, Abbas Karami |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
DNA-programmable assembly
02 engineering and technology 010402 general chemistry 01 natural sciences Article law.invention Post-PCR detection chemistry.chemical_compound law Spherical nucleic acids (SNAs) Materials Chemistry Electrical and Electronic Engineering Instrumentation Gene Polymerase Polymerase chain reaction ComputingMethodologies_COMPUTERGRAPHICS biology Metals and Alloys RNA Amplicon 021001 nanoscience & nanotechnology Condensed Matter Physics Molecular biology 0104 chemical sciences Surfaces Coatings and Films Electronic Optical and Magnetic Materials Coronavirus disease 2019 (COVID-19) chemistry biology.protein Nucleic acid 5’-exonuclease activity 0210 nano-technology Linker DNA Colorimetric detection |
| Zdroj: | Sensors and Actuators. B, Chemical Sensors and Actuators B: Chemical |
| ISSN: | 0925-4005 |
| Popis: | Graphical abstract Highlights • The amplification power of PCR is coupled with programmability of SNA assembly for detection of COVID-19. • A linker-based single-component assembly system based on E gene of SARS-CoV-2 RNA provides a post PCR colorimetric response with comparable sensitivity to real time RT-PCR. • The palindromic linker is disintegrated based on 5’-exonuclease activity in the course of PCR amplification. • The positive and negative viral COVID-19 samples are simply distinguished via different colors by naked eyes. Continuous identification of suspected infectious cases is crucial to control the recent pandemic caused by the novel human coronavirus SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2). Real-time polymerase chain reaction (real-time PCR) technology cannot be implemented easily and in large scale in some communities due to lack of resources and infrastructures. Here, we report a simple colorimetric strategy derived from linker-based single-component assembly of gold nanoparticle core-spherical nucleic acids (AuNP-coreSNAs) for visual detection of PCR products of SARS-CoV-2 ribonucleic acid (RNA) template. A palindromic linker is designed based on E gene of SARS-CoV-2 to program the identical colloidal SNAs into large assemblies along with a distinct red-to-purple color change. The linker acts as a probe of SARS-CoV-2 RNA in conventional PCR reaction. In the presence of the correct template, the palindromic linker, which is complementary to a short region within the amplicon, is cleaved by 5’-exonuclease activity of deoxyribonucleic acid (DNA) polymerase. Disintegrating the palindromic linker during the amplification process inhibits the post single-component assembly formation of SNAs. So, positive and negative viral samples produce simply red and purple colors in the post PCR colorimetric test, respectively. Evaluation of the samples obtained from cases with laboratory-confirmed SARS-CoV-2 infection revealed that our assay can rival with Real-time PCR method in sensitivity. |
| Databáze: | OpenAIRE |
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