Development of a LAMP assay with a portable device for real-time detection of begomoviruses under field conditions
| Autor: | Yutaro Neriya, Tomohide Natsuaki, Sedyo Hartono, Hiroshi Katoh, Fariha Wilisiani, Aika Tomiyama, Hisashi Nishigawa |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Veterinary medicine Time Factors Melon 030106 microbiology Loop-mediated isothermal amplification 03 medical and health sciences Virology Fluorometer Pepper Fluorometry Solanaceae DNA Primers Plant Diseases biology fungi Begomovirus food and beverages biology.organism_classification Plant Leaves Cucurbitaceae 030104 developmental biology Indonesia Nucleic Acid Amplification Techniques Field conditions |
| Zdroj: | Journal of virological methods. 265 |
| ISSN: | 1879-0984 |
| Popis: | The emergence of begomovirus infection is one of the most important problems affecting production of a variety of vegetable crops worldwide. Infection by begomoviruses has been detected and spread rapidly on Cucurbitaceae and Solanaceae plants in Indonesia. A rapid and simple detection assay for begomoviruses under field conditions for routine sampling of plants is needed. Primers for a loop-mediated isothermal amplification (LAMP) assay were designed based on the sequences of three Indonesian begomoviruses, namely Tomato leaf curl New Delhi virus (ToLCNDV), Pepper yellow leaf curl Indonesia virus (PepYLCIV), and Tomato yellow leaf curl Kanchanaburi virus (TYLCKaV), infecting Cucurbitaceae and Solanaceae plants. LAMP assays using a Genelyzer™ III portable fluorometer with a toothpick method successfully detected these begomoviruses in infected melon, pepper, and eggplant samples. LAMP assays conducted during a field survey for detection of the three begomoviruses on 104 fresh leaves indicated that most of the samples were positive; the findings were confirmed by PCR using universal primers of begomovirus as a common detection method. These results demonstrate that this simple and rapid LAMP assay using a fluorometer portable device may be used to achieve real-time detection of begomoviruses under field conditions. |
| Databáze: | OpenAIRE |
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