@article{10902/32719,
year = {2024},
month = {4},
url = {https://hdl.handle.net/10902/32719},
abstract = {Gene sequencing in back of reverse transcription-quantitative polymerase chain reaction (RT-qPCR) is the current approach for discriminating infections produced by different severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants in the clinic. However, sequencing is often a time-consuming step, which hinders the deployment of a very fast response during a pandemic. Here, we propose to run a CRISPR-Cas12a reaction after completing the RT-qPCR and in the very same pot to detect with high specificity genetic marks characterizing variants of concern. A crRNA was appropriately designed to detect the S gene of the SARS-CoV-2 Omicron BA.1 variant. A significant response with >20-fold dynamic range was obtained for the Omicron BA.1 S gene, while the Delta S gene did not produce any detectable signal. The sensitivity of the method was analyzed with a series of diluted samples and different Cas12a nucleases. A correlation between the RT-qPCR CT values and the CRISPR-Cas12a reaction signals was observed. Variant discrimination with the CRISPR-Cas12a reaction was possible in some minutes with high accuracy from patient samples. In conclusion, CRISPR-Cas systems seem ready to be exploited in the clinic to boost personalized diagnoses and accelerate epidemiological surveillance in a cost-effective way.},
organization = {Work supported by Generalitat Valenciana (GVA-COVID19/2021/036), Ministerio de Ciencia e Innovación (PDC2022−133941-I00; cofunded by NextGenerationEU), CRUE and Banco Santander (Fondo Supera Covid-19), CSIC PTI Salud Global (NextGenerationEU Fund, regulation 2020/2094), and IDIVAL (INNVAL21/13).},
publisher = {American Chemical Society},
publisher = {ACS Omega 2024, 9, 18046-18050},
title = {Rapid and accurate detection of the SARS-CoV-2 omicron variant with a CRISPR-Cas12a reaction in the RT-qPCR Pot},
author = {Rodrigo, Guilermo and Ruiz González, Raúl and Montagud Martínez, Roser and Dorta Gorrín, Alexis and Pablo Marcos, Daniel and Gozalo Margüello, Mónica and Calvo Montes, Jorge and Navas Méndez, Jesús},
}