Comparison and application of RT-LAMP and RT-PCR methods for MCMV detection

For the detection of Maize chlorotic mottle virus（MCMV），a reverse-transcription，loop mediated isothermal amplification（ RT-LAMP） assay with high sensitivity and specificity was established. A set of five primers were designed，based on the coat-protein sequence of MCMV. By optimizing the concentration of primers，the best temperature，the suitable reaction time，and loop primer，the RT-LAMP was built by incubation at 64℃ for only 45 min with loop primer. The detection limit of RT-LAMP assay was 280 fg of MCMV RNA，which was 100 times more sensitive than that of RT- PCR assay. High species-specificity of RT- LAMP method was confirmed by the assay of 5 pathogens such as MCMV，SCMV，MDMV， WSMV and MWLMV. In addition，all the simulated samples were detected by LAMP assay，detection rate was 100%.