Evaluation of a lyophilized CRISPR-Cas12 assay for a sensitive, specific, and rapid detection of SARS-CoV-2
- Autores
- Curti, Lucía Ana; Primost, Ivana; Valla, Sofia; Ibañez Alegre, Daiana; Olguin Perglione, Cecilia; Repizo, Guillermo Daniel; Lara, Julia; Parcerisa, Ivana; Palacios, Antonela; Llases, María Eugenia; Rinflerch, Adriana; Barrios, Melanie; Pereyra Bonnet, Federico; Gimenez, Carla Alejandra; Marcone, Débora Natalia
- Año de publicación
- 2021
- Idioma
- inglés
- Tipo de recurso
- artículo
- Estado
- versión publicada
- Descripción
- We evaluated a lyophilized CRISPR-Cas12 assay for SARS-CoV-2 detection (Lyo-CRISPR SARS-CoV-2 kit) based on reverse transcription, isothermal amplification, and CRISPR-Cas12 reaction. From a total of 210 RNA samples extracted from nasopharyngeal swabs using spin columns, the Lyo-CRISPR SARS-CoV-2 kit detected 105/105 (100%; 95% confidence interval (CI): 96.55–100) positive samples and 104/105 (99.05%; 95% CI: 94.81–99.97) negative samples that were previously tested using commercial RT-qPCR. The estimated overall Kappa index was 0.991, reflecting an almost perfect concordance level between the two diagnostic tests. An initial validation test was also performed on 30 nasopharyngeal samples collected in lysis buffer, in which the Lyo-CRISPR SARS-CoV-2 kit detected 20/21 (95.24%; 95% CI: 76.18–99.88) positive samples and 9/9 (100%; 95% CI: 66.37–100) negative samples. The estimated Kappa index was 0.923, indicating a strong concordance between the test procedures. The Lyo-CRISPR SARS-CoV-2 kit was suitable for detecting a wide range of RT-qPCR-positive samples (cycle threshold range: 11.45–36.90) and dilutions of heat-inactivated virus (range: 2.5–100 copies/µL); no cross-reaction was observed with the other respiratory pathogens tested. We demonstrated that the performance of the Lyo-CRISPR SARS-CoV-2 kit was similar to that of commercial RT-qPCR, as the former was highly sensitive and specific, timesaving (1.5 h), inexpensive, and did not require sophisticated equipment. The use of this kit would reduce the time taken for diagnosis and facilitate molecular diagnosis in low-resource laboratories.
Instituto de Virología
Fil: Curti, Lucía Ana. CASPR Biotech; Estados Unidos
Fil: Primost, Ivana. Hospital Municipal de Trauma y Emergencias Dr. Federico Abete. Genetics and Molecular Biology Laboratory; Argentina
Fil: Valla, Sofia. Universidad Nacional del Noroeste de la Provincia de Buenos Aires. Centro de Investigaciones y Transferencia del Noroeste de la Provincia de Buenos Aires (CITNOBA). Centro de Investigaciones Básicas y Aplicadas (CIBA); Argentina
Fil: Valla, Sofia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Ibañez Alegre, Daiana. Universidad Nacional de Misiones. Instituto de Biología Subtropical. Laboratorio Grupo de Investigación en Genética Aplicada (GIGA); Argentina
Fil: Ibañez Alegre, Daiana. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Olguin Perglione, Cecilia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina
Fil: Olguin Perglione, Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Repizo, Guillermo Daniel. CASPR Biotech; Estados Unidos
Fil: Lara, Julia. CASPR Biotech; Estados Unidos
Fil: Parcerisa, Ivana. CASPR Biotech; Estados Unidos
Fil: Palacios, Antonela. CASPR Biotech; Estados Unidos
Fil: Llases, María Eugenia. CASPR Biotech; Estados Unidos
Fil: Rinflerch, Adriana. Universidad Nacional de Misiones. Instituto de Biología Subtropical. Laboratorio Grupo de Investigación en Genética Aplicada (GIGA); Argentina
Fil: Rinflerch, Adriana. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Barrios, Melanie. Universidad de Buenos Aires. Instituto de Producción Agropecuaria; Argentina
Fil: Pereyra Bonnet, Federico. CASPR Biotech; Estados Unidos
Fil: Gimenez, Carla Alejandra. CASPR Biotech; Estados Unidos
Fil: Marcone, Débora Natalia. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología, Biotecnología y Genética. Cátedra de Virología; Argentina
Fil: Marcone, Débora Natalia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina - Fuente
- Viruses 13 (3) : 420 (Marzo 2021)
- Materia
-
CRISPR
Severe Acute Respiratory Syndrome Coronavirus 2
COVID-19
Diagnosis
Fluorescence
Freeze Drying
Repeticiones Palindrómicas Cortas Agrupadas y Regularmente Interespaciadas
Coronavirus del Síndrome Respiratorio Agudo Grave 2
Diagnóstico
Fluorescencia
Liofilización
SARS-CoV-2 - Nivel de accesibilidad
- acceso abierto
- Condiciones de uso
- http://creativecommons.org/licenses/by-nc-sa/4.0/
- Repositorio
.jpg)
- Institución
- Instituto Nacional de Tecnología Agropecuaria
- OAI Identificador
- oai:localhost:20.500.12123/9128
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Evaluation of a lyophilized CRISPR-Cas12 assay for a sensitive, specific, and rapid detection of SARS-CoV-2Curti, Lucía AnaPrimost, IvanaValla, SofiaIbañez Alegre, DaianaOlguin Perglione, CeciliaRepizo, Guillermo DanielLara, JuliaParcerisa, IvanaPalacios, AntonelaLlases, María EugeniaRinflerch, AdrianaBarrios, MelaniePereyra Bonnet, FedericoGimenez, Carla AlejandraMarcone, Débora NataliaCRISPRSevere Acute Respiratory Syndrome Coronavirus 2COVID-19DiagnosisFluorescenceFreeze DryingRepeticiones Palindrómicas Cortas Agrupadas y Regularmente InterespaciadasCoronavirus del Síndrome Respiratorio Agudo Grave 2DiagnósticoFluorescenciaLiofilizaciónSARS-CoV-2We evaluated a lyophilized CRISPR-Cas12 assay for SARS-CoV-2 detection (Lyo-CRISPR SARS-CoV-2 kit) based on reverse transcription, isothermal amplification, and CRISPR-Cas12 reaction. From a total of 210 RNA samples extracted from nasopharyngeal swabs using spin columns, the Lyo-CRISPR SARS-CoV-2 kit detected 105/105 (100%; 95% confidence interval (CI): 96.55–100) positive samples and 104/105 (99.05%; 95% CI: 94.81–99.97) negative samples that were previously tested using commercial RT-qPCR. The estimated overall Kappa index was 0.991, reflecting an almost perfect concordance level between the two diagnostic tests. An initial validation test was also performed on 30 nasopharyngeal samples collected in lysis buffer, in which the Lyo-CRISPR SARS-CoV-2 kit detected 20/21 (95.24%; 95% CI: 76.18–99.88) positive samples and 9/9 (100%; 95% CI: 66.37–100) negative samples. The estimated Kappa index was 0.923, indicating a strong concordance between the test procedures. The Lyo-CRISPR SARS-CoV-2 kit was suitable for detecting a wide range of RT-qPCR-positive samples (cycle threshold range: 11.45–36.90) and dilutions of heat-inactivated virus (range: 2.5–100 copies/µL); no cross-reaction was observed with the other respiratory pathogens tested. We demonstrated that the performance of the Lyo-CRISPR SARS-CoV-2 kit was similar to that of commercial RT-qPCR, as the former was highly sensitive and specific, timesaving (1.5 h), inexpensive, and did not require sophisticated equipment. The use of this kit would reduce the time taken for diagnosis and facilitate molecular diagnosis in low-resource laboratories.Instituto de VirologíaFil: Curti, Lucía Ana. CASPR Biotech; Estados UnidosFil: Primost, Ivana. Hospital Municipal de Trauma y Emergencias Dr. Federico Abete. Genetics and Molecular Biology Laboratory; ArgentinaFil: Valla, Sofia. Universidad Nacional del Noroeste de la Provincia de Buenos Aires. Centro de Investigaciones y Transferencia del Noroeste de la Provincia de Buenos Aires (CITNOBA). Centro de Investigaciones Básicas y Aplicadas (CIBA); ArgentinaFil: Valla, Sofia. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Ibañez Alegre, Daiana. Universidad Nacional de Misiones. Instituto de Biología Subtropical. Laboratorio Grupo de Investigación en Genética Aplicada (GIGA); ArgentinaFil: Ibañez Alegre, Daiana. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Olguin Perglione, Cecilia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; ArgentinaFil: Olguin Perglione, Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Repizo, Guillermo Daniel. CASPR Biotech; Estados UnidosFil: Lara, Julia. CASPR Biotech; Estados UnidosFil: Parcerisa, Ivana. CASPR Biotech; Estados UnidosFil: Palacios, Antonela. CASPR Biotech; Estados UnidosFil: Llases, María Eugenia. CASPR Biotech; Estados UnidosFil: Rinflerch, Adriana. Universidad Nacional de Misiones. Instituto de Biología Subtropical. Laboratorio Grupo de Investigación en Genética Aplicada (GIGA); ArgentinaFil: Rinflerch, Adriana. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Barrios, Melanie. Universidad de Buenos Aires. Instituto de Producción Agropecuaria; ArgentinaFil: Pereyra Bonnet, Federico. CASPR Biotech; Estados UnidosFil: Gimenez, Carla Alejandra. CASPR Biotech; Estados UnidosFil: Marcone, Débora Natalia. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología, Biotecnología y Genética. Cátedra de Virología; ArgentinaFil: Marcone, Débora Natalia. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaMDPI2021-04-19T17:40:04Z2021-04-19T17:40:04Z2021-03info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttp://hdl.handle.net/20.500.12123/9128https://www.mdpi.com/1999-4915/13/3/4201999-4915https://doi.org/10.3390/v13030420Viruses 13 (3) : 420 (Marzo 2021)reponame:INTA Digital (INTA)instname:Instituto Nacional de Tecnología Agropecuariaenginfo:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by-nc-sa/4.0/Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)2025-09-29T13:45:11Zoai:localhost:20.500.12123/9128instacron:INTAInstitucionalhttp://repositorio.inta.gob.ar/Organismo científico-tecnológicoNo correspondehttp://repositorio.inta.gob.ar/oai/requesttripaldi.nicolas@inta.gob.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:l2025-09-29 13:45:11.824INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuariafalse |
| dc.title.none.fl_str_mv |
Evaluation of a lyophilized CRISPR-Cas12 assay for a sensitive, specific, and rapid detection of SARS-CoV-2 |
| title |
Evaluation of a lyophilized CRISPR-Cas12 assay for a sensitive, specific, and rapid detection of SARS-CoV-2 |
| spellingShingle |
Evaluation of a lyophilized CRISPR-Cas12 assay for a sensitive, specific, and rapid detection of SARS-CoV-2 Curti, Lucía Ana CRISPR Severe Acute Respiratory Syndrome Coronavirus 2 COVID-19 Diagnosis Fluorescence Freeze Drying Repeticiones Palindrómicas Cortas Agrupadas y Regularmente Interespaciadas Coronavirus del Síndrome Respiratorio Agudo Grave 2 Diagnóstico Fluorescencia Liofilización SARS-CoV-2 |
| title_short |
Evaluation of a lyophilized CRISPR-Cas12 assay for a sensitive, specific, and rapid detection of SARS-CoV-2 |
| title_full |
Evaluation of a lyophilized CRISPR-Cas12 assay for a sensitive, specific, and rapid detection of SARS-CoV-2 |
| title_fullStr |
Evaluation of a lyophilized CRISPR-Cas12 assay for a sensitive, specific, and rapid detection of SARS-CoV-2 |
| title_full_unstemmed |
Evaluation of a lyophilized CRISPR-Cas12 assay for a sensitive, specific, and rapid detection of SARS-CoV-2 |
| title_sort |
Evaluation of a lyophilized CRISPR-Cas12 assay for a sensitive, specific, and rapid detection of SARS-CoV-2 |
| dc.creator.none.fl_str_mv |
Curti, Lucía Ana Primost, Ivana Valla, Sofia Ibañez Alegre, Daiana Olguin Perglione, Cecilia Repizo, Guillermo Daniel Lara, Julia Parcerisa, Ivana Palacios, Antonela Llases, María Eugenia Rinflerch, Adriana Barrios, Melanie Pereyra Bonnet, Federico Gimenez, Carla Alejandra Marcone, Débora Natalia |
| author |
Curti, Lucía Ana |
| author_facet |
Curti, Lucía Ana Primost, Ivana Valla, Sofia Ibañez Alegre, Daiana Olguin Perglione, Cecilia Repizo, Guillermo Daniel Lara, Julia Parcerisa, Ivana Palacios, Antonela Llases, María Eugenia Rinflerch, Adriana Barrios, Melanie Pereyra Bonnet, Federico Gimenez, Carla Alejandra Marcone, Débora Natalia |
| author_role |
author |
| author2 |
Primost, Ivana Valla, Sofia Ibañez Alegre, Daiana Olguin Perglione, Cecilia Repizo, Guillermo Daniel Lara, Julia Parcerisa, Ivana Palacios, Antonela Llases, María Eugenia Rinflerch, Adriana Barrios, Melanie Pereyra Bonnet, Federico Gimenez, Carla Alejandra Marcone, Débora Natalia |
| author2_role |
author author author author author author author author author author author author author author |
| dc.subject.none.fl_str_mv |
CRISPR Severe Acute Respiratory Syndrome Coronavirus 2 COVID-19 Diagnosis Fluorescence Freeze Drying Repeticiones Palindrómicas Cortas Agrupadas y Regularmente Interespaciadas Coronavirus del Síndrome Respiratorio Agudo Grave 2 Diagnóstico Fluorescencia Liofilización SARS-CoV-2 |
| topic |
CRISPR Severe Acute Respiratory Syndrome Coronavirus 2 COVID-19 Diagnosis Fluorescence Freeze Drying Repeticiones Palindrómicas Cortas Agrupadas y Regularmente Interespaciadas Coronavirus del Síndrome Respiratorio Agudo Grave 2 Diagnóstico Fluorescencia Liofilización SARS-CoV-2 |
| dc.description.none.fl_txt_mv |
We evaluated a lyophilized CRISPR-Cas12 assay for SARS-CoV-2 detection (Lyo-CRISPR SARS-CoV-2 kit) based on reverse transcription, isothermal amplification, and CRISPR-Cas12 reaction. From a total of 210 RNA samples extracted from nasopharyngeal swabs using spin columns, the Lyo-CRISPR SARS-CoV-2 kit detected 105/105 (100%; 95% confidence interval (CI): 96.55–100) positive samples and 104/105 (99.05%; 95% CI: 94.81–99.97) negative samples that were previously tested using commercial RT-qPCR. The estimated overall Kappa index was 0.991, reflecting an almost perfect concordance level between the two diagnostic tests. An initial validation test was also performed on 30 nasopharyngeal samples collected in lysis buffer, in which the Lyo-CRISPR SARS-CoV-2 kit detected 20/21 (95.24%; 95% CI: 76.18–99.88) positive samples and 9/9 (100%; 95% CI: 66.37–100) negative samples. The estimated Kappa index was 0.923, indicating a strong concordance between the test procedures. The Lyo-CRISPR SARS-CoV-2 kit was suitable for detecting a wide range of RT-qPCR-positive samples (cycle threshold range: 11.45–36.90) and dilutions of heat-inactivated virus (range: 2.5–100 copies/µL); no cross-reaction was observed with the other respiratory pathogens tested. We demonstrated that the performance of the Lyo-CRISPR SARS-CoV-2 kit was similar to that of commercial RT-qPCR, as the former was highly sensitive and specific, timesaving (1.5 h), inexpensive, and did not require sophisticated equipment. The use of this kit would reduce the time taken for diagnosis and facilitate molecular diagnosis in low-resource laboratories. Instituto de Virología Fil: Curti, Lucía Ana. CASPR Biotech; Estados Unidos Fil: Primost, Ivana. Hospital Municipal de Trauma y Emergencias Dr. Federico Abete. Genetics and Molecular Biology Laboratory; Argentina Fil: Valla, Sofia. Universidad Nacional del Noroeste de la Provincia de Buenos Aires. Centro de Investigaciones y Transferencia del Noroeste de la Provincia de Buenos Aires (CITNOBA). Centro de Investigaciones Básicas y Aplicadas (CIBA); Argentina Fil: Valla, Sofia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Ibañez Alegre, Daiana. Universidad Nacional de Misiones. Instituto de Biología Subtropical. Laboratorio Grupo de Investigación en Genética Aplicada (GIGA); Argentina Fil: Ibañez Alegre, Daiana. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Olguin Perglione, Cecilia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina Fil: Olguin Perglione, Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Repizo, Guillermo Daniel. CASPR Biotech; Estados Unidos Fil: Lara, Julia. CASPR Biotech; Estados Unidos Fil: Parcerisa, Ivana. CASPR Biotech; Estados Unidos Fil: Palacios, Antonela. CASPR Biotech; Estados Unidos Fil: Llases, María Eugenia. CASPR Biotech; Estados Unidos Fil: Rinflerch, Adriana. Universidad Nacional de Misiones. Instituto de Biología Subtropical. Laboratorio Grupo de Investigación en Genética Aplicada (GIGA); Argentina Fil: Rinflerch, Adriana. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Barrios, Melanie. Universidad de Buenos Aires. Instituto de Producción Agropecuaria; Argentina Fil: Pereyra Bonnet, Federico. CASPR Biotech; Estados Unidos Fil: Gimenez, Carla Alejandra. CASPR Biotech; Estados Unidos Fil: Marcone, Débora Natalia. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología, Biotecnología y Genética. Cátedra de Virología; Argentina Fil: Marcone, Débora Natalia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina |
| description |
We evaluated a lyophilized CRISPR-Cas12 assay for SARS-CoV-2 detection (Lyo-CRISPR SARS-CoV-2 kit) based on reverse transcription, isothermal amplification, and CRISPR-Cas12 reaction. From a total of 210 RNA samples extracted from nasopharyngeal swabs using spin columns, the Lyo-CRISPR SARS-CoV-2 kit detected 105/105 (100%; 95% confidence interval (CI): 96.55–100) positive samples and 104/105 (99.05%; 95% CI: 94.81–99.97) negative samples that were previously tested using commercial RT-qPCR. The estimated overall Kappa index was 0.991, reflecting an almost perfect concordance level between the two diagnostic tests. An initial validation test was also performed on 30 nasopharyngeal samples collected in lysis buffer, in which the Lyo-CRISPR SARS-CoV-2 kit detected 20/21 (95.24%; 95% CI: 76.18–99.88) positive samples and 9/9 (100%; 95% CI: 66.37–100) negative samples. The estimated Kappa index was 0.923, indicating a strong concordance between the test procedures. The Lyo-CRISPR SARS-CoV-2 kit was suitable for detecting a wide range of RT-qPCR-positive samples (cycle threshold range: 11.45–36.90) and dilutions of heat-inactivated virus (range: 2.5–100 copies/µL); no cross-reaction was observed with the other respiratory pathogens tested. We demonstrated that the performance of the Lyo-CRISPR SARS-CoV-2 kit was similar to that of commercial RT-qPCR, as the former was highly sensitive and specific, timesaving (1.5 h), inexpensive, and did not require sophisticated equipment. The use of this kit would reduce the time taken for diagnosis and facilitate molecular diagnosis in low-resource laboratories. |
| publishDate |
2021 |
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2021-04-19T17:40:04Z 2021-04-19T17:40:04Z 2021-03 |
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article |
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http://hdl.handle.net/20.500.12123/9128 https://www.mdpi.com/1999-4915/13/3/420 1999-4915 https://doi.org/10.3390/v13030420 |
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http://hdl.handle.net/20.500.12123/9128 https://www.mdpi.com/1999-4915/13/3/420 https://doi.org/10.3390/v13030420 |
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1999-4915 |
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eng |
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MDPI |
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Viruses 13 (3) : 420 (Marzo 2021) reponame:INTA Digital (INTA) instname:Instituto Nacional de Tecnología Agropecuaria |
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