Evaluation of a lyophilized crispr-cas12 assay for a sensitive, specific, and rapid detection of sars-cov-2

Autor: Cecilia Olguin Perglione, Julia Lara, Carla Alejandra Gimenez, Daiana Macarena Ibáñez Alegre, Melanie Barrios, Federico Alberto Pereyra Bonnet, Ivana Parcerisa, Lucia Ana Curti, Antonela Palacios, Ivana Primost, Adriana Raquel Rinflerch, Débora Natalia Marcone, Guillermo Daniel Repizo, Sofia Valla, María Eugenia Llases
Jazyk: angličtina
Rok vydání: 2021
Předmět:
0301 basic medicine
Serial dilution
diagnosis
Repeticiones Palindrómicas Cortas Agrupadas y Regularmente Interespaciadas
viruses
LYSIS BUFFER
lcsh:QR1-502
lcsh:Microbiology
purl.org/becyt/ford/1 [https]
0302 clinical medicine
COVID-19 Testing
Nasopharynx
Clustered Regularly Interspaced Short Palindromic Repeats
030212 general & internal medicine
skin and connective tissue diseases
Chemistry
Diagnóstico
Severe Acute Respiratory Syndrome Coronavirus 2
Respiratory pathogens
Infectious Diseases
Molecular Diagnostic Techniques
CRISPR
Fluorescencia
RNA
Viral
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)
Loop-mediated isothermal amplification
SARS-COV-2
DIAGNOSIS
Rapid detection
Sensitivity and Specificity
Article
Fluorescence
lysis buffer
Liofilización
03 medical and health sciences
Virology
Lysis buffer
Humans
purl.org/becyt/ford/1.6 [https]
Cycle threshold
Chromatography
FLUORESCENCE DETECTION
SARS-CoV-2
fungi
COVID-19
Highly sensitive
respiratory tract diseases
body regions
030104 developmental biology
Freeze Drying
fluorescence detection
Coronavirus del Síndrome Respiratorio Agudo Grave 2
Zdroj: CONICET Digital (CONICET)
Consejo Nacional de Investigaciones Científicas y Técnicas
instacron:CONICET
Viruses
Volume 13
Issue 3
Viruses, Vol 13, Iss 420, p 420 (2021)
Viruses 13 (3) : 420 (Marzo 2021)
INTA Digital (INTA)
Instituto Nacional de Tecnología Agropecuaria
instacron:INTA
Popis: 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.
Databáze: OpenAIRE
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