Reverse Transcription-loop-mediated Isothermal Amplification for Detection of Maize Chlorotic Mottle Virus
Maize chlorotic mottle virus (MCMV) is a quarantine pest as approved by Chinese government.A rapid,sensitive and specific MCMV detection method using reverse transcription-loop-mediated isothermal amplification (RT-LAMP) was established in this study.Based on the sequence of MCMV coat protein coding gene,3 sets of primers were designed and specificity test showed that the second set of primers was specific to MCMV.Similar sensitivities were observed on RT-LAMP and RT-PCR,except that RT-LAMP was quicker,and the reaction could be finished within 1 h.In addition,the presence or absence of the fluorescence under daylight allows naked easy detection of the amplification of MCMV genomic RNA using calcein.The RT-LAMP assay was applied successfully to detect MCMV in maize seeds,and the result by the addition of calcein was consistent with the result detected by the real time turbidimeter.The method is rapid,specific,sensitive without the need for complicated equipment,and is suitable for rapid field detection of MCMV.
NETL
NSTL
万方数据
维普
