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 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.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2023 |
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| Erschienen: |
2023 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:12 |
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| Enthalten in: |
Biology - 12(2023), 7 vom: 23. Juni |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
García-Sorribes, Soraya [VerfasserIn] |
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| Links: |
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| Themen: |
COVID-19 |
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| Anmerkungen: |
Date Revised 31.07.2023 published: Electronic Citation Status PubMed-not-MEDLINE |
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| doi: |
10.3390/biology12070900 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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NLM360082033 |
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| 520 | |a 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 | ||
| 650 | 4 | |a Journal Article | |
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| 650 | 4 | |a SARS-CoV-2 | |
| 650 | 4 | |a colorimetry | |
| 650 | 4 | |a trypsin | |
| 700 | 1 | |a Lara-Hernández, Francisco |e verfasserin |4 aut | |
| 700 | 1 | |a Manzano-Blasco, Iris |e verfasserin |4 aut | |
| 700 | 1 | |a Abadía-Otero, Jessica |e verfasserin |4 aut | |
| 700 | 1 | |a Albert, Eliseo |e verfasserin |4 aut | |
| 700 | 1 | |a Mulet, Alba |e verfasserin |4 aut | |
| 700 | 1 | |a Briongos-Figuero, Laisa Socorro |e verfasserin |4 aut | |
| 700 | 1 | |a Gabella-Martín, Miriam |e verfasserin |4 aut | |
| 700 | 1 | |a Torres, Ignacio |e verfasserin |4 aut | |
| 700 | 1 | |a Signes-Costa, Jaime |e verfasserin |4 aut | |
| 700 | 1 | |a Navarro, David |e verfasserin |4 aut | |
| 700 | 1 | |a Martín-Escudero, Juan-Carlos |e verfasserin |4 aut | |
| 700 | 1 | |a García-García, Ana-Bárbara |e verfasserin |4 aut | |
| 700 | 1 | |a Chaves, Felipe Javier |e verfasserin |4 aut | |
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