Sample Treatment with Trypsin for RT-LAMP COVID-19 Diagnosis

The SARS-CoV-2 coronavirus is responsible for the COVID-19 pandemic resulting in a global health emergency. Given its rapid spread and high number of infected individuals, a diagnostic tool for a rapid, simple, and cost-effective detection was essential. In this work, we developed a COVID-19 diagnos...

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Published in:Biology (Basel, Switzerland) Switzerland), 2023-06, Vol.12 (7), p.900
Main Authors: García-Sorribes, Soraya, Lara-Hernández, Francisco, Manzano-Blasco, Iris, Abadía-Otero, Jessica, Albert, Eliseo, Mulet, Alba, Briongos-Figuero, Laisa Socorro, Gabella-Martín, Miriam, Torres, Ignacio, Signes-Costa, Jaime, Navarro, David, Martín-Escudero, Juan-Carlos, García-García, Ana-Bárbara, Chaves, Felipe Javier
Format: Article
Language:English
Subjects:
Brief Report
Colorimetry
Coronaviruses
COVID-19
Diagnosis
Ethylenediaminetetraacetic acid
Exudates
Gene amplification
Genetic transcription
Health aspects
Laboratories
Pandemics
Pharmaceutical industry
Public health
Reverse transcription
Ribonucleic acid
RNA
RNA extraction-free
RT-LAMP
Saliva
SARS-CoV-2
Sensitivity analysis
Severe acute respiratory syndrome coronavirus 2
Temperature
Trypsin
Viruses
World health
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Summary:The SARS-CoV-2 coronavirus is responsible for the COVID-19 pandemic resulting in a global health emergency. Given its rapid spread and high number of infected individuals, a diagnostic tool for a rapid, simple, and cost-effective detection was essential. In this work, we developed a COVID-19 diagnostic test, that incorporates a human internal control, based on the Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP). When working with synthetic SARS-CoV-2 RNA, the optimized RT-LAMP assay has a sensitivity of 10 viral copies and can be detected by fluorescence in less than 15 min or by the naked eye in 25 min using colorimetric RT-LAMP. To avoid the RNA extraction step, a pre-treatment of the sample was optimized. Subsequently, a validation was performed on 268 trypsin treated samples (including nasopharyngeal, buccal, and nasal exudates) and amplified with colorimetric RT-LAMP to evaluate its sensitivity and specificity in comparison with RT-qPCR of extracted samples. The validation results showed a sensitivity and specificity of 100% for samples with Ct ≤ 30. The rapid, simple, and inexpensive RT-LAMP SARS-CoV-2 extraction-free procedure developed may be an alternative test that could be applied for the detection of SARS-CoV-2 or adapted to detect other viruses present in saliva or nasopharyngeal samples with higher sensitivity and specificity of the antibody test.
ISSN:2079-7737
2079-7737
DOI:10.3390/biology12070900