Fuente: PubMed "stone fruits" J Virol Methods. 2026 Jun 10;345:115429. doi: 10.1016/j.jviromet.2026.115429. Online ahead of print.ABSTRACTHigh-throughput sequencing has expanded the known diversity of viruses infecting stone fruits within the family Betaflexiviridae, particularly the subfamily Quinvirinae, complicating routine diagnostics based on species-specific primers. In this study, a universal, degenerate primer set targeting conserved regions of the RNA-dependent RNA polymerase (RdRp) was designed by aligning public sequences from Robigovirus and Foveavirus (4-5 isolates per species where available) and evaluated to enable broad, non-specific detection of multiple Betaflexiviridae species in stone fruits and selected non-Prunus hosts. The assay was validated on (i) reference sources for CNRMV, CGRMV, CRMaV, PVGR, WCaV, ApLV; (ii) locally collected Prunus samples (n = 449); and (iii) non-Prunus hosts, pear (ASPV) and grapevine (GRSPaV). Universal RT-qPCR screening (SYBR Green), followed by conventional RT-PCR and Sanger sequencing (∼280 nt), enabled species attribution and confirmation with species-specific primers. Among 449 Prunus accessions, 45 (10.0%) were qPCR-positive and yielded amplicons of the expected size by RT-PCR. Prevalence was highest in sweet cherry (20.0%, 30/150), followed by apricot (13.3%, 8/60) and peach (5.4%, 7/130); no infection was detected in plum. BLAST analysis identified 21 Robigovirus positives (13 PVGR in cherry; 8 CGRMV in cherry, peach, apricot), 4 Foveavirus (ApLV in apricot and peach; APV2 in apricot and peach), 19 CVA (Capillovirus; 14 in cherry and 5 in apricot/peach), and 1 AVCaV (Prunevirus; apricot). In addition, ASPV was detected in pear and GRSPaV in grapevine. This RdRp-based primer set provides broad detection across Quinvirinae and selected Trivirinae, streamlining large-scale screening and first-line diagnostics for stone fruit health programs. Species-level resolution is readily achieved by Sanger sequencing or secondary species-specific RT-PCR, supporting application in surveillance, certification, and quarantine.PMID:42263865 | DOI:10.1016/j.jviromet.2026.115429