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严格的种子检疫是控制病原体跨区域远距离传播的最有效方法之一。瓜类细菌性果斑病、黄瓜细菌性角斑病、黄瓜绿斑驳花叶病毒病和黄瓜黑星病均是严重危害瓜类作物的重要病害。本研究基于微流控和实时荧光定量PCR(Real-time quantitative PCR,qPCR)技术，结合核酸提取纯化、多目标快速扩增和结果可视化技术，对上述瓜类作物重要病原物进行多重检测。整个检测过程仅需36 min，瓜类细菌性果斑病菌最低检测限为2.1×10~3CFU/mL，黄瓜细菌性角斑病菌最低检测限为2.4×10~3CFU/mL，黄瓜绿斑驳花叶病毒最低检测限为1.0×10~1copies/μL，黄瓜黑星病菌最低检测限为1.7×10-3ng/μL。使用该方法检测了47份瓜类实际样品，与实验室q PCR方法的检测结果一致。该方法摆脱了对实验室和专业技术人员的依赖，有助于提高植物有害生物的现场检测技术水平。
Abstract：Strict seed quarantine is one of the most effective ways to control the long-distance spread of pathogens across regions. Bacterial fruit blotch of cucurbits,cucumber angular leaf spot,cucumber green mottle mosaic virus disease and scab of cucurbits are the important diseases of cucurbits. In this study,based on microfluidic and real-time quantitative PCR(qPCR),we combined nucleic acid extraction and purification,multi-target rapid amplification and result visualization for multiple targets detection of pathogens in cucurbita crops. The entire detection process only took 36 minutes. The limit of detection(LOD) were 2.1×103 CFU/mL,2.4×103 CFU/mL,1.0×101 copies/μL and 1.7×10-3ng/μL for Acidovorax citrulli,Pseudomonas syringae pv. lachrymans,cucumber green mottle mosaic virus and Cladosporium cucumerinum,respectively. This method was used to detect 47 samples,which were consistent with the test results obtained by the q PCR. This method helps to eliminate the dependence on laboratories and professional technicians,and improves the technical level of on-site detection of plant pathogens.
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基本信息:
DOI：10.19662/j.cnki.issn1005-2755.2025.06.004
中图分类号:S41-30
引用信息:
[1]王梦雨,蔡璐璐,孙羽佳,等.瓜类作物重要病原物多目标现场快检方法的研究与应用[J].植物检疫,2025,39(06):19-27.DOI:10.19662/j.cnki.issn1005-2755.2025.06.004.
基金信息:
中国检验检疫科学研究院基本科研业务费项目（2024JK016）; 国家市场监督管理总局科技创新人才计划（KJLJ202317）