Development of an RT-LAMP assay for the detection of maize yellow mosaic virus in maize
| Autor: | Hu Wenli, Xiaoqin Li, Suiyun Chen, Jianguang Wang, Yan Li, Yu Li |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
Detection limit
food.ingredient Loop-mediated isothermal amplification RNA Reverse Transcription Biology Molecular biology Sensitivity and Specificity Zea mays Maize yellow mosaic virus Polerovirus Luteoviridae food Real-time polymerase chain reaction Molecular Diagnostic Techniques Mosaic Viruses Virology Nucleic Acid Amplification Techniques Family Luteoviridae Plant Diseases |
| Zdroj: | Journal of virological methods. 300 |
| ISSN: | 1879-0984 |
| Popis: | Maize is one of the most widely cultivated cereal crops worldwide. Maize yellow mosaic virus (MaYMV) (species Maize yellow mosaic virus, genus Polerovirus and family Luteoviridae) was first reported in maize from China. In this study, a one-step reverse-transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for detecting MaYMV. The optimal concentrations of betaine, Mg2+ and dNTPs for the assay were 0 M, 1.4 mM and 6 mM, respectively, and the optimal reaction time was 50 min. Using total plant RNA as the template, the detection limit of the RT-LAMP assay for MaYMV was 1 pg, while that of RT-PCR was 100 pg, indicating that the RT-LAMP assay developed was 100 times more sensitive than RT-PCR. Importantly, the RT-LAMP assay successfully detected MaYMV using rapidly extracted crude RNA from infected maize as a template. In conclusion, the RT-LAMP assay developed was a rapid, specific, sensitive and low-cost method for the detection of MaYMV in field samples of maize. |
| Databáze: | OpenAIRE |
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