Publications
BiologyJun 2023 |
12
(
7
),
900
DOI:
10.3390/biology12070900

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

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
Product Used
NGS
Abstract
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.
Product Used
NGS

Related Publications