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/μL. 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.

登革热病毒（DENV）是威胁人类生命的病毒之一。在这项研究中，我们设计了特定的新型引物，用于通过实时逆转录环介导的等温扩增（RT-LAMP）反应快速判别检测DENV-1，DENV-2和DENV-4。研究了反应温度和硫酸镁等参数对RT-LAMP反应检测DENV RNA的影响。在最佳条件下，该方法能够在25分钟内区分和检测DENV，检测极限为3.5拷贝/μL。重要的是，新的基于特异性引物的RT-LAMP测定法不与其他病毒反应，表明该方法对DENV RNA的选择性。将RT-LAMP反应产物在365 nm的紫外光下照射时，很容易用肉眼看到。放大后的产品可以直接看到颜色变化。该方法为快速鉴别检测登革热病毒提供了一种简便，准确的分子扩增技术。RT-LAMP平台可用作区分性检测DENV的重要诊断工具，而无需使用复杂的协议或复杂的设备。