Development of simultaneous detection method of six viruses infecting grapevine by a multiplex RT-PCR assay

Abstract A multiplex RT-PCR is quick and economical method because of the simultaneous detection for two or more targets. Here, we developed multiplex RT-PCR for six grapevine viruses. Primers were newly designed for each viral isolate based on complete sequence data obtained from the NCBI database. As the result of comparing the detection sensitivities of our multiplex RT-PCR assay with those of previously reported conventional singleplex RT-PCR assays, that of our assay exhibit equivalent or even 1–10,000 times greater than their assays without non-specific reactions, respectively. Our assay is useful in selecting healthy nurseries and are useful for plant quarantine inspections.