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Rapid and Reliable Detection of SARS-CoV-2 Using Direct RT-LAMP
- Source :
- Diagnostics, Vol 12, Iss 4, p 828 (2022)
- Publication Year :
- 2022
- Publisher :
- MDPI AG, 2022.
-
Abstract
- Background: The global pandemic coronavirus SARS-CoV-2 has a healthcare, social and economic burden. To limit the spread of the virus, the World Health Organization (WHO) urgently called for extensive screening of suspected individuals; thus, a quick, simple, and sensitive diagnostic assay is always in need. Methods: We applied reverse transcription-loop-mediated isothermal amplification (RT-LAMP) for the detection of SARS-CoV-2. The RT-LAMP method was optimized by evaluating two fluorescence amplification mixes and several reaction times, and results were compared to the standard real-time RT-PCR (rtRT-PCR). The assay was validated using 200 nasopharyngeal swabs collected in viral transport media (62 positive for SARS-CoV-2, and 138 negative for SARS-CoV-2 detected by the rtRT-PCR method). The samples were diluted 1:4 in diethylpyrocarbonate (DEPC)-treated water, utilized for RT-LAMP using different singleplex and multiplex sets of LAMP primers (N gene, S gene, and orf1ab gene), and incubated at 65 °C using real-time PCR 7500. Results: Our direct detection with the RT-LAMP protocol showed 100% concordance (sensitivity and specificity) with the standard protocol used for the detection of SARS-CoV-2 nucleic acid. Conclusions: In this study, we set up a rapid, simple, and sensitive RT-LAMP assay for the detection of SARS-CoV-2 in clinical samples. The assay is suitable for point of care detection in public hospitals, medical centers in rural areas, and in transportation hubs.
- Subjects :
- RT-LAMP
direct assay
SARS-CoV-2
COVID-19
molecular assay
PCR
Medicine (General)
R5-920
Subjects
Details
- Language :
- English
- ISSN :
- 20754418
- Volume :
- 12
- Issue :
- 4
- Database :
- Directory of Open Access Journals
- Journal :
- Diagnostics
- Publication Type :
- Academic Journal
- Accession number :
- edsdoj.42c2f0d1667e4f94a17a5dd945910c2f
- Document Type :
- article
- Full Text :
- https://doi.org/10.3390/diagnostics12040828