| Autor: |
García-Sorribes S; Genomic and Diabetes Unit, INCLIVA Biomedical Research Institute, 46010 Valencia, Spain., Lara-Hernández F; Genomic and Diabetes Unit, INCLIVA Biomedical Research Institute, 46010 Valencia, Spain., Manzano-Blasco I; Genomic and Diabetes Unit, INCLIVA Biomedical Research Institute, 46010 Valencia, Spain., Abadía-Otero J; Internal Medicine Service, Rio Hortega University Hospital, 47012 Valladolid, Spain., Albert E; Microbiology Service, University Clinic Hospital, INCLIVA, 46010 Valencia, Spain., Mulet A; Pulmonary Department, University Clinic Hospital, INCLIVA, 46010 Valencia, Spain., Briongos-Figuero LS; Internal Medicine Service, Rio Hortega University Hospital, 47012 Valladolid, Spain., Gabella-Martín M; Internal Medicine Service, Rio Hortega University Hospital, 47012 Valladolid, Spain., Torres I; Microbiology Service, University Clinic Hospital, INCLIVA, 46010 Valencia, Spain., Signes-Costa J; Pulmonary Department, University Clinic Hospital, INCLIVA, 46010 Valencia, Spain., Navarro D; Microbiology Service, University Clinic Hospital, INCLIVA, 46010 Valencia, Spain., Martín-Escudero JC; Internal Medicine Service, Rio Hortega University Hospital, 47012 Valladolid, Spain.; Medicine Department, Valladolid University, 47002 Valladolid, Spain., García-García AB; Genomic and Diabetes Unit, INCLIVA Biomedical Research Institute, 46010 Valencia, Spain.; CIBERDEM, ISCIII, 28029 Madrid, Spain., Chaves FJ; Genomic and Diabetes Unit, INCLIVA Biomedical Research Institute, 46010 Valencia, Spain.; CIBERDEM, ISCIII, 28029 Madrid, Spain. |
| Abstrakt: |
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. |
