A Rapid and Highly Sensitive Assay for the detection of Dahlia mosaic caulimovirus in Dahlia (Dahlia variabilis)
Primary author: Romana Iftikhar
Authors: Romana Iftikhar, Lindani Moyo, Hanu Pappu
Primary college/unit: Agricultural, Human and Natural Resource Sciences
Dahlia (Dahlia variabilis) is a highly valued ornamental plant widely grown in the U.S. and several other countries for its flowers, and the Pacific Northwest boasts more than a dozen dahlia societies. Dahlia mosaic virus (DMV), an aphid-transmitted caulimovirus in the family Caulimoviridae, is an important pathogen of dahlia (Dahlia variabilis). DMV genome consists of a circular, double-stranded DNA, approximately 8kb in size, and the DNA is packaged in spherical virus particles. Development of rapid, and sensitive diagnostic tools is essential in surveillance and management of DMV.
A loop-mediated isothermal amplification (LAMP) assay was developed for DMV detection. The LAMP assay targets the reverse transcriptase region of the DMV genome and is optimal at a temperature of 60°C and run time of 60 min, and amplification was detected through fluorescence detection and by agarose gel electrophoresis. The assay detected DMV in DNA concentration as low as 10 ? 3 ng. The LAMP assay was found to be more sensitive than polymerase chain reaction test. To the best of our knowledge, this is the first report of a LAMP assay for the detection of DMV, a plant DNA virus. This assay will be useful in rapid and sensitive detection of DMV and in reducing the virus incidence through production of virus-free planting material.