期刊
TALANTA
卷 190, 期 -, 页码 391-396出版社
ELSEVIER
DOI: 10.1016/j.talanta.2018.08.019
关键词
Dengue virus; Specific primers; Aedes aegypi mosquito; Reverse trascription loop-mediated isothermal amplification; Molecular diagnosis
资金
- National Research Foundation (NRF) - Ministry of Science and ICT, South Korea [NRF-2016M3A9B6939558]
- Chung-Ang University
Dengue virus (DENV) is one of the life-threatening viruses to the human. In this study, we have designed specific novel primers for rapid discriminative detection of DENV-1, DENV-2, and DENV-4 by real-time reverse transcription loop-mediated isothermal amplification (RT-LAMP) reaction. The effect of parameters such as reaction temperature and magnesium sulfate was investigated on the RT-LAMP reaction for detection of DENV RNA. Under the optimal conditions, this method is able to differentiate and to detect DENV within 25 min, exhibiting detection limit of 3.5 copies/mu L. Importantly, the novel specific primers-based RT-LAMP assay did not react with other viruses, suggesting the selectivity of the method towards DENV RNA. The RT-LAMP reaction products are easily visualized with naked-eye when irradiated them under UV light at 365 nm. Amplification products could be visualized directly for color changes. This method provides a facile, and accurate molecular amplication technique for the rapid discriminative detection of dengue viruses. The RT-LAMP platform can be used as a promissing diagnostic tool for discriminative detection of DENV without aid of complicated protocols or sophisticated equipment.
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