ArticleFrontiers in cellular and infection microbiology2021
Rapid Detection of SARS-CoV-2 Using Duplex Reverse Transcription-Multienzyme Isothermal Rapid Amplification in a Point-of-Care Testing.
Article in Frontiers in cellular and infection microbiology, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed.
- Development of a MIRA-CRISPR/Cas12a-based nucleic acid detection system for the discrimination ofJournal of ginseng research · 2026Article
- Advances in rapid detection technologies for zoonotic diseases: a one health-oriented review.Frontiers in cellular and infection microbiology · 2026Review
- Evaluation and Application of the MIRA-qPCR Method for Rapid Detection of Norovirus Genogroup II in Shellfish.Microorganisms · 2025Article
- Real-Time Reverse Transcription Multienzyme Isothermal Rapid Amplification for Rapid Detection of African Horse Sickness Virus.Transboundary and emerging diseases · 2025Article
- A novel system with robust compatibility and stability for detectingMicrobiology spectrum · 2024Article
- A novel detection method based on MIRA-CRISPR/Cas13a-LFD targeting the repeated DNA sequence of Trichomonas vaginalis.Parasites & vectors · 2024Article
- Rapid detection of human cytomegalovirus by multienzyme isothermal rapid amplification and lateral flow dipsticks.Frontiers in cellular and infection microbiology · 2024Article
- Review
- A fully integrated duplex RT-LAMP device for the detection of viral infections.Biomedical microdevices · 2023Article
- Development and Evaluation of Duplex MIRA-qPCR Assay for Simultaneous Detection of Staphylococcus aureus and non-aureus StaphylococciMicroorganisms · 2022Article
- Rapid detection of SARS-CoV-2: The gradual boom of lateral flow immunoassay.Frontiers in bioengineering and biotechnology · 2022Review
- Rapid detection ofFrontiers in microbiology · 2022Article
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Authors and funding
9 authors.
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Abstract
In December 2019, a severe acute respiratory syndrome caused by SARS-CoV-2 spread rapidly worldwide. Portable nucleic acid tests of SARS-CoV-2 are critically important for diagnostics. In this study, we used an isothermal amplification method-Multienzyme Isothermal Rapid Amplification (MIRA)-for rapid detection of SARS-CoV-2. We designed the primers and probes in ORF1ab and N gene of SARS-CoV-2. The amplicons could be monitored by lateral flow dipsticks (LFDs). The reaction temperature, time, concentrations of primers and probes, and working volume were optimized. Four commercial swab collection buffers were used to test the amplification efficacy of our assay without RNA extraction. Our assay was able to amplify duplex targets of SARS-CoV-2 in one single reaction using one-step RT-MIRA. The assay worked well in a low volume of 10 μl at 38°C for 20 min. Using three collection buffers without guanidinium, our assay was able to amplify efficaciously without RNA extraction. The 95% limit of detection (LoD) of the RT-MIRA assay was 49.5 (95% CI, 46.8-52.7) copies/ml for ORF1ab gene and 48.8 (95% CI, 46.5-52.6) copies/ml for N gene. There is no cross-reaction with other human respiratory pathogens, such as SARS-CoV, MERS-CoV, influenza A virus, influenza B virus, human adenovirus, respiratory syncytial virus, human parainfluenza virus, and coronavirus 229E in our assay. The precision evaluation revealed that the C50
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