Establishment of a triplex TaqMan quantitative real-time PCR assay for simultaneous detection of Cymbidium mosaic virus, Odontoglossum ringspot virus and Cymbidium ringspot virus

• DOI: 10.3389/fmicb.2023.1129259
• Publication Date: 2023-05-19T05:13:36Z
• Authors: Aiqing Sun; Lihua Wang; Yiping Zhang; Xiumei Yang; Yi Wei; Dong Yang; Wenhan Li; Xuewei Wu
• Keywords: TaqMan

Orchids are significant ornamental plants whose viral infection results in substantial economic damage. Cymbidium mosaic virus (CymMV), Odontoglossum ringspot (ORSV), and (CymRSV) represent three important prevalent orchid viruses. The detection system proposed this study uses a triplex TaqMan quantitative real-time PCR assay to identify CymMV, ORSV, CymRSV simultaneous manner. We designed specific primers probes for CymRSV, with amplified sequences of 156 bp, 148 145 respectively. minimum limit the qRT-PCR CymMV was 1 copy/assay, 10 copies/assay ORSV. stable limits were 10, 102, 102 copies/assay, Therefore, exhibited higher sensitivity (approximately 104-fold) than RT-PCR. intra-and interassay CVs Cq values less 0.55 0.95%, respectively, indicating that is highly reliable accurate. In addition, 66 samples from five different genera analyzed using established gene chip. demonstrated probe chip, could be used field samples. Our findings suggest RT-PCR represents rapid, simple, accurate tool detecting on orchids.