A CRISPR-Cas12a-based diagnostic method for multiple genotypes of severe fever with thrombocytopenia syndrome virus
Fig 2
Detection of lentiviruses pseudotyped with SFTSV S gene using DETECTR combined with fluorescence assay.
(A) A diagram of gRNAs and primer sets for SFTSV DETECTR. (B and C) Lentiviruses pseudotyped with and without SFTSV S gene (SPV and CPV, respectively) (100 CFU), IAV and IBV (100 PFU) and HCV and HEV (100 genome copies) were lysed via HUDSON and viral nucleic acids amplified via RT-RPA with a primer set specific for the SFTSV S gene. RT-RPA amplicons were detected with SFTSV DETECTR combined with fluorescence assay using gRNAs (A) S1 and (B) S2 corresponding to the S genes of all SFTSV genotypes. Fluorescence saturation occurred within 10 min. Values are presented as means ± s.d. (error bars) (n = 3 replicates; * p < 0.05 between samples, two-sample t-test). nt, nucleotides; PFU, plaque forming unit; CFU, colony forming unit; SPV, SFTSV pseudovirus; CPV, control pseudovirus.