College of Plant Protection, Shandong Agricultural University, Taian, Shandong, China.
Facility Horticulture Laboratory of Universities in Shandong, Weifang University of Science and Technology, Shouguang, Shandong, China.
PLoS One. 2020 Mar 5;15(3):e0230023. doi: 10.1371/journal.pone.0230023. eCollection 2020.
Melon necrotic spot virus (MNSV) can cause significant economic losses due to decreased quality in cucurbit crops. The current study is the first to use reverse transcription loop-mediated isothermal amplification (RT-LAMP) for detection of MNSV. A set of four LAMP primers was designed based on the coat protein gene sequence of MNSV, and a RT-LAMP reaction was successfully performed for 1 h at 62°C. The results of RT-LAMP showed high specificity for MNSV and no cross-reaction with other viruses. Compared to traditional reverse transcription-PCR (RT-PCR), the RT-LAMP assay was 103-fold more sensitive in detecting MNSV. Due to its sensitivity, speed and visual assessment, RT-LAMP is appropriate for detecting MNSV in the laboratory.
蜜瓜果斑坏死病毒（MNSV）可导致葫芦科作物品质下降，造成重大经济损失。本研究首次采用逆转录环介导等温扩增（RT-LAMP）技术检测 MNSV。根据 MNSV 外壳蛋白基因序列设计了 4 条 LAMP 引物，并在 62°C 下成功进行了 1 小时的 RT-LAMP 反应。RT-LAMP 结果显示对 MNSV 具有高度特异性，与其他病毒无交叉反应。与传统的逆转录聚合酶链反应（RT-PCR）相比，RT-LAMP 检测 MNSV 的灵敏度高 103 倍。由于其灵敏度、速度和可视化评估，RT-LAMP 适用于实验室检测 MNSV。
