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 Rapid discriminative detection of Dengue viruses via loop mediated isothermal amplification
Tác giả hoặc Nhóm tác giả: Jong-Gil Kim, Seung Hoon Baek, Seungrok Kim, Hae In Kim, Seung Woo Lee, Le Minh Tu Phan, Suresh Kumar Kailasa, and Tae Jung Park
Nơi đăng: Talanta; Số: 190;Từ->đến trang: 391-396;Năm: 2018
Lĩnh vực: Y - Dược; Loại: Bài báo khoa học; Thể loại: Quốc tế
TÓM TẮT
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, suggestng 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 techique 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.
ABSTRACT
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, suggestng 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 techique 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.
[ 2018\2018m08d014_18_25_11Talanta-Rapid_discriminative_detection_of_Dengue_viruses_via_loop_mediated_isothermal_amplification.pdf ]
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