Development of reverse transcription loop-mediated isothermal amplification assay for sensitive and rapid detection of Hosta virus X

The one-step real-time turbidity loop-mediated isothermal amplification assay (RealAmp) was developed to detect Hosta virus X (HVX), the most devastating threat to hosta industry. The reaction was performed in a single tube at 63 degrees C for 15min, and real-time turbidimetry was used to monitor the amplification results. Specificity and sensitivity analyses demonstrated that this RealAmp method was sensitive as real-time TaqMan RT-PCR and about 100-fold higher than conventional RT-PCR with no cross-reaction with other viral pathogens. Field samples detection showed that HVX could be identified effectively with this method. Overall, this RealAmp assay for HVX detection was simple, specific, sensitive, convenient and time-saving and could assist in the quarantine measures for prevention and control of the disease caused by HVX.