基于环介导等温扩增的石蒜绵粉蚧可视化快速检测OA
Visual and rapid detection of Phenacoccus solani based on loop-mediated isothermal amplification
[目的]石蒜绵粉蚧是一种世界性花果害虫,为及时、有效控制其危害,开展该虫的可视化快速检测.[方法]以石蒜绵粉蚧的28S rDNA序列为靶标,设计其特异性的环介导等温扩增(loop-mediated isothermal amplifi-cation,LAMP)引物组,用于建立相应的可视化快速检测方法,并进行特异性、灵敏度及稳定性验证.[结果]建立的LAMP可视化快速检测方法能有效扩增石蒜绵粉蚧DNA,最佳反应温度和时间及最低检测浓度分别为64℃、60 min和100 fg/µL,反应产物呈蓝色阳性,检出率达95.83%～100.00%;该方法无法扩增近缘种属粉蚧DNA,反应产物呈紫色阴性.[结论]石蒜绵粉蚧的LAMP可视化快速检测方法特异、灵敏、稳定,可用于该虫的现场识别.
[Objective]Phenacoccus solani is a global pest of fruits and flowers.It is necessary to developed a visual and rapid detection method for the control of this mealybug.[Method]Based on the 28S rDNA sequence of P.solani,the specific primers of loop-mediated isothermal amplification(LAMP)were designed to establish the corresponding visual detection method,and the specificity,sensitivity and stability of LAMP detection were verified.[Result]The LAMP detection at 64℃for 60 min effectively detected the DNA of P.solani,it had a detection limit of 100 fg/µL and the detection ratio of 95.83%～100.00%with the positive blue reaction products.And the LAMP detection could not amplify the DNA of other mealybug species with the negative purple reaction products.[Conclusion]The LAMP visual and rapid detection is specific,sensitive and reliable for identification of P.solani on-site.
张杰;蓝炎阳;姚锦爱;余德亿;黄鹏
福建省农业科学院植物保护研究所,福建 福州 350013漳州市花卉研究所,福建 漳州 363902福建省农业科学院植物保护研究所,福建 福州 350013福建省农业科学院植物保护研究所,福建 福州 350013福建省农业科学院植物保护研究所,福建 福州 350013
农业科技
石蒜绵粉蚧环介导等温扩增可视化快速检测现场识别
Phenacoccus solaniloop-mediated isothermal amplificationvisual and rapid detectionon-site identification
《东南园艺》 2026 (1)
67-72,6
福建省自然科学基金项目(2024J01324)福建省属公益类科研院所基本科研专项(2024R1023004)
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