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- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 2020.1
- 수록면
- 1 - 15 (15page)
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SARS-CoV-2 is very contagious and has rapidly spread globally. Due to various symptomatic and asymptomatic cases and the possibility of asymptomatic transmission, there is a pressing need for a fast and sensitive detection protocol to diagnose asymptomatic people. Various SARS-CoV-2 diagnostic kits are already available from many companies and national health agencies. However, publicly available information on these diagnostic kits is lacking. In response to the growing need and the lack of information, we developed and made available a low-cost, easy-access, real-time PCRbased protocol for the early detection of the virus in a previous study. During the development of the detection protocol, we found that unoptimized primer sets could inadvertently show false-positive results, raising the possibility that commercially available diagnostic kits might also contain primer sets that produce false-positive results. Here, we provide three-step guidelines for the design and optimization of specific primer sets. The three steps include (1) the selection of primer sets for target genes (RdRP, N, E, and S) in the genome of interest (SARS-CoV-2), (2) the in silico validation of primer and amplicon sequences, and (3) the optimization of PCR conditions (i.e., primer concentrations and annealing temperatures) for specific hybridization between the primers and target genes, and the elimination of spurious primer dimers. Furthermore, we have expanded the previously developed real-time PCR-based protocol to more conventional PCR-based protocols and applied a multiplex PCR-based protocol that allows the simultaneous testing of primer sets for RdRP, N, E, and S all in one reaction. Our newly optimized protocol should be helpful for the large-scale, high-fidelity screening of asymptomatic people, even without any high-specification equipment, for the further prevention of transmission, and to achieve early intervention and treatment for the rapidly propagating virus.
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POCT Detection of?14 Respiratory Viruses Using Multiplex RT-PCR
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Development of Universal Primers for Use in RT-qPCR and RT-RPA Targeting SARS-CoV-2 Genes
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Development of Various Diagnostic tool of SARS-COV-2 Infection
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Novel Method to Reduce Non-specific Reaction in Real-time Recombinase Polymerase Amplification Using Primers Containing an AP site for Multiplex Detection of SARS CoV-2
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Phylogenetic Relationship of the Complete SARS-CoV-2 Genome Sequences
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SARS-CoV-2 Recovery from the Wastewater
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Development of PCR Primers for Carbonic Anhydrase Gene of Genus Bradyrhizobium
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2018 .10
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On the origin and evolution of SARS-CoV-2
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Discrimination and isolation of the virus from free RNA fragments for the highly sensitive measurement of SARS-CoV-2 abundance on surfaces using a graphene oxide nano surface
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