可可花瘿病菌LAMP检测方法的建立
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1福州海关技术中心,福建福州 350001 ; 2. 福建农林大学植物保护学院,福建福州 350002 ; 3. 漳州市植保站,福建漳州 363000

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李敏女高级农艺师?研究方向:植物病害检疫及检测技术E-mail:limin_2926@163.com

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福建省中央引导地方科技发展专项(2022L3016)


Development of a LAMP assay for rapid detection of Albonectria rigidiuscula
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1 Technology Center of Fuzhou Customs District, Fuzhou, Fujian 350001 , China ; 2. College of Plant Protection, Fujian Agriculture and Forestry University, Fuzhou, Fujian 350002 , China ; 3. Zhangzhou Plant Protection Station, Zhangzhou, Fujian 363000 , China

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    摘要:

    【目的】建立可可花瘿病菌快速、灵敏、便捷、可视的环介导等温扩增技术(loop-mediated isothermal amplification,LAMP)检测方法,为口岸外来有害生物的快速检测、精准防控和保护我国生态安全提供技术支持。【方法】以可可花瘿病菌EF-1α和Tub基因序列为靶序列,设计LAMP引物并筛选出有效引物1组,优化反应温度,测试其特异性和灵敏度,并对该病菌的寄主进行实际检测。【结果】本研究建立的可视化LAMP检测方法可在1h内对可可花瘿病菌进行检测,供试其他19株病原菌均未出现扩增,仅可可花瘿病菌出现扩增,具有良好的特异性;检测温度为65℃,检测灵敏度可达21.84×10-4 ng·μL-1,且在实际应用中准确率高达100%;检测结果可直接肉眼判断。【结论】本研究建立的可可花瘿病菌LAMP检测方法不仅反应时间短、特异性强、灵敏度高、操作简单,且检测结果可视化,适用于可可花瘿病菌的口岸检疫及田间检测。

    Abstract:

    【Aim】This study aims to develop a rapid, sensitive, convenient, and visual loop-mediated isothermal amplification (LAMP) detection method for Albonectria rigidiuscula, providing technical support for the rapid detection and accurate management of this invasive pest.【Method】LAMP primers were designed based on the EF-1α and Tub sequences of A. rigidiuscula, and a set of effective primers was screened. The reaction temperature was optimized, and the specificity and sensitivity of the method were evaluated. Additionally, the host of A. rigidiuscula was detected.【Result】The visual LAMP detection method established in this study achieved the detection of A. rigidiuscula within 1 h. No amplification band was produced for the 19 other pathogenic strains tested, and only A. rigidiuscula exhibited amplification, which demonstrated the excellent specificity of the established method. The optimal detection temperature was 65 ℃, and the detection sensitivity reached 21.84×10-4 ng·μL-1. In practical applications, the method achieved an accuracy rate of 100%, and the test results could be intuitively interpreted.【Conclusion】The LAMP method developed in this study for the detection of A. rigidiuscula features a short reaction time, strong specificity, high sensitivity, simple operation, and visual detection results. It is suitable for port quarantine and field detection of A. rigidiuscula.

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李敏,牟茜,潘永红,胡美玲,虞赟,沈建国.可可花瘿病菌LAMP检测方法的建立[J].生物安全学报中文版,2026,35(2):195-202

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  • 收稿日期:2025-04-19
  • 最后修改日期:2025-08-18
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  • 在线发布日期: 2026-06-03
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