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Little cherry virus 1 (LChV-1, Velarivirus, Closteroviridae) is an economically important pathogen affect-ing mainly cherry around the world emphasizing the impetus for its efficient and accurate on-site de-tection. This study describes the development of a reliable diagnostic protocol of LChV-1 based on a fast, sensitive, and easy-to-use one-step reverse-transcription loop-mediated isothermal amplification (RT-LAMP). The protocol detects all LChV-1 isolates in less than 10 min by fluorescence monitoring using a mobile detection device (GENIE, OptiGene) and is most optimal when incubation is performed at 67 °C. Sharp melting curves and unique melting temperatures (Tm) were obtained for the positive samples. Both the LAMP and classical RT-PCR methods are capable of specifically detecting LChV-1 in infected leaf tissues, but this validated leaf-to-result assay has remarkable advantages in comparison to RT-PCR. It is at least hundred fold more sensitive, significantly faster, and efficient at minimal cost, and is ready for on-field applications. In conclusion, this innovative LAMP approach can contribute to implementation of sustainable integrated management strategies for on-site detection of LChV-1 in commercial orchards or for horticultural research stations. It is also suitable for decision support in phytosanitary epidemiological programs.
Original languageEnglish
Title of host publicationNew and fast detection of Little cherry virus 1 using one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP)
Number of pages1
Publication date22-May-2018
Publication statusPublished - 22-May-2018
Event61st International Symposium on Crop Protection (2009): ISCP - FBW, UGent, Gent, Belgium
Duration: 22-May-201822-May-2018

ID: 6225020