Desenho e validação in silico de iniciadores para detecção do coronavírus 2 causador da síndrome respiratória aguda grave (Sars-Cov-2)

Abstract

The design of polymerase chain reaction (PCR) primers that target conserved segmentsofviral genomes is important to prevent false-negative results and reduce the need tostandardizedifferent PCR protocols for the same target. In this work, we designed and describedaset ofprimers and probes that target conserved regions identified from multiple alignment of 2,341SARS-CoV-2 genomes available in the GISAID (Global Initiative on Sharing All InfluenzaData) database. Subsequently, the primers were validated together with the probes on211,833sequences from the entire genomes of SARS-CoV-2. Nine systems were obtained(primerforward+reverse+probes) that potentially anneal to the highly conserved regions of thevirusgenome identified in this analysis. In silico predictions also demonstrated that the primersdonot interact with non-specific targets in sequences from humans, bacteria, fungi, Apicomplexaand other betacoronaviruses and less pathogenic coronavirus strains. The publicationof theseprimer and probes sequences will make it possible to validate more efficient protocolsforidentifying SARS-CoV-2.