ArticleDiagnostics (Basel, Switzerland)2022
A Novel Rolling Circle Amplification-Based Detection of SARS-CoV-2 with Multi-Region Padlock Hybridization.
Article in Diagnostics (Basel, Switzerland), 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 8 papers.
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Who cites it
8 citing papers in PubMed.
- Indiscriminate Trans-Cleavage Activity of CRISPR/SuCas12a2 Enables Sensitive Detection of SARS-CoV-2.ACS sensors · 2026Article
- Establishment of a Padlock Probe-Based Fluorescence Quantitative PCR Method for the Detection of the SARS-CoV-2 Omicron S371L Mutation.Journal of fluorescence · 2026Article
- Circular nucleic acids at the host-virus interface: from immune modulation to therapeutic innovation.Frontiers in immunology · 2026Review
- Next-Generation Nucleic Acid-Based Diagnostics for Viral Pathogens: Lessons Learned from the SARS-CoV-2 Pandemic.Microorganisms · 2025Review
- Enhancing Clinical Detection Accuracy of Large Structured Viral RNA via DNAzyme Cleavage and Antisense-Assisted Rolling Circle Amplification.Angewandte Chemie (International ed. in English) · 2025Article
- Optimization and Benchmarking of RT-LAMP-CRISPR-Cas12a for the Detection of SARS-CoV-2 in Saliva.International journal of molecular sciences · 2025Article
- Nucleic Acid Based Testing (NABing): A Game Changer Technology for Public Health.Molecular biotechnology · 2024Review
- ARTseq-FISH reveals position-dependent differences in gene expression of micropatterned mESCs.Nature communications · 2024Article
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Authors and funding
11 authors.
Funding
Abstract
SARS-CoV-2 has remained a global health burden, primarily due to the continuous evolution of different mutant strains. These mutations present challenges to the detection of the virus, as the target genes of qPCR, the standard diagnostic method, may possess sequence alterations. In this study, we develop an isothermal one-step detection method using rolling circle amplification (RCA) for SARS-CoV-2. This novel strategy utilizes a multi-padlock (MP-RCA) approach to detect viral-RNA via a simplified procedure with the reliable detection of mutated strains over other procedures. We designed 40 padlock-based probes to target different sequences across the SARS-CoV-2 genome. We established an optimal one-step isothermal reaction protocol utilizing a fluorescent output detected via a plate reader to test a variety of padlock combinations. This method was tested on RNA samples collected from nasal swabs and validated via PCR. S-gene target failure (SGTF)-mutated strains of SARS-CoV-2 were included. We demonstrated that the sensitivity of our assay was linearly proportional to the number of padlock probes used. With the 40-padlock combination the MP-RCA assay was able to correctly detect 45 out 55 positive samples (81.8% efficiency). This included 10 samples with SGTF mutations which we were able to detect as positive with 100% efficiency. We found that the MP-RCA approach improves the sensitivity of the MP-RCA assay, and critically, allows for the detection of SARS-CoV-2 variants with SGTF. Our method offers the simplicity of the reaction and requires basic equipment compared to standard qPCR. This method provides an alternative approach to overcome the challenges of detecting SARS-CoV-2 and other rapidly mutating viruses.
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