期刊
ACTA VIROLOGICA
卷 58, 期 2, 页码 160-166出版社
AEPRESS SRO
DOI: 10.4149/av_2014_02_160
关键词
beet curly top virus; Curtovirus; detection; Geminivirus; loop-mediated isothermal amplification
类别
资金
- Animal, Plant & Fisheries Quarantine and Inspection Agency, Ministry of Food, Agriculture, Forestry and Fisheries, Republic of Korea [Z-1541785-2012-12-03]
- iPET (Korea Institute of Planning and Evaluation for Technology in Food, Agriculture, Forestry and Fisheries), Ministry for Agriculture, Food and Rural Affairs, Republic of Korea [311058-05-3-HD140]
Rapid and sensitive detection methods for three species of Curtovirus were developed using a loop-mediated isothermal amplification (LAMP) technique. A universal primer set for detecting the three main species of Curtovirus at the same time, and three kinds of species-specific primer sets were designed and used for LAMP reactions. Results from the LAMP reactions were visualized both by color changes after adding SYBR Green I staining dye and by DNA laddering on agarose gel electrophoresis. The optimal conditions for the curtovirus LAMP reaction were confirmed at 60 degrees C for the universal primers and at 62 degrees C for the three species-specific primer sets. Amplification of curtoviruses by LAMP reaction was ten-fold more sensitive than that by polymerase chain reaction. Primers designed for curtovirus detection in this study did not anneal to or amplify DNA from other DNA or RNA viruses (tomato yellow leaf curl virus, tomato spotted wilt virus, and potato virus Y). Taken together, the primer sets and reaction conditions developed in this study show that the LAMP technique could be a useful tool to detect the three species of Curtovirus simultaneously and distinguish them in the laboratory and the field.
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