Development of real-time reverse transcription PCR for detection of Maize chlorotic mottle virus based on a novel molecular marker

Maize chlorotic mottle virus (MCMV) infects maize plants and causes significant losses in corn production worldwide. In this study, a real-time TaqMan RT-PCR assay for efficient detection of MCMV was described. A pair of primers amplifying a 131-bp DNA fragment and a TaqMan probe was designed targeting the novel molecular marker based on MCMV genome analysis sequences. The assay designed was highly specific, producing no signal from other viruses, and the sensitivity of the assay was 0.16 fg/reaction of total RNA, which was approximately 252-fold higher than conventional RT-PCR gel electrophoresis method. Compared with the real-time TaqMan reverse transcription PCR targeting coat protein gene, the novel assay has more specificity and sensitivity to detect MCMV in maize. Therefore, the assay is a useful tool for large or middle-scale corporations and entry-exit inspection and quarantine bureau to detect MCMV in maize seeds or plant tissues