Simultaneous detection of four lily-infecting viruses by a multiplex RT-PCR assay

A multiplex reverse-transcription polymerase chain reaction (RT-PCR) system was modified and evaluated for the simultaneous detection of multiple viruses in coinfected lily plants. Four major lily viruses, namely, lily symptomless, cucumber mosaic, lily mottle, and plantago asiatica mosaic viruses, were targeted by simultaneous detection. Analysis of PCR products confirmed the amplification of virus-specific DNA fragments of the expected sizes and nucleotide sequences from the target viruses. The cDNAs from singly infected plant samples were used as positive controls, and multiple infected samples were used to develop the multiplex RT-PCR system, which was then applied to leaf samples from diverse lily species and hybrids. The modified multiplex RT-PCR was successfully validated on the simultaneous detection of these viruses.