基于TaqMan探针实时荧光定量PCR的白蜡鞘孢菌分子鉴定技术OA
Molecular identification technology of Hymenoscyphus fraxineus based on TaqMan qPCR
为快速准确检测白蜡枯梢病病原菌白蜡鞘孢菌Hymenoscyphus fraxineus,选择ITS和翻译延长因子1α(TEF1)基因为靶标,设计特异性引物与探针,构建目标菌株的TaqMan探针qPCR检测体系,并通过优化退火温度和引物与探针浓度完善该检测体系.结果表明,TaqMan探针qPCR体系可特异性扩增目标菌株靶标基因,最低DNA检测浓度为1.0 pg/μL;利用该检测体系可实现对白蜡鞘孢菌的高效率鉴定和疫情监控,为口岸外来有害生物防控、生态安全保护提供技术服务与支持.
To rapidly and accurately detect Hymenoscyphus fraxineus,pathogenic fungus of ash dieback,internal transcribed spacer(ITS)and translation elongation factor 1-α(TEF1)genes were selected as targets for designing specific primers and TaqMan probes.A TaqMan probe-based qPCR detection system was established for target strains,and assay was optimized by adjusting annealing temperatures and concentrations of primers and probes.The results showed that TaqMan qPCR system could specifically amplify target genes of target strain,and minimum detection concentration of DNA was 1.0 pg/μL.The detection system could enable efficient quarantine identification and epidemic monitoring of H.fraxineus,and provide technical services and support for prevention and control of invasive pests at ports and protection of ecological security.
杨春光;李辉;刘亚乔;薛腾;李鑫
大连海关技术中心,辽宁 大连 116001东港海关综合技术服务中心,辽宁 丹东 118399大窑湾海关综合技术服务中心,辽宁 大连 116100中华人民共和国大连邮局海关,辽宁 大连 116014大连海关技术中心,辽宁 大连 116001
农业科技
TaqMan探针实时荧光定量PCR白蜡鞘孢菌灵敏度
TaqMan probereal-time fluorescence quantitative PCRHymenoscyphus fraxineussensitivity
《中国森林病虫》 2026 (1)
60-64,5
国家重点研发计划项目"外来病虫害高效检测关键技术与装备研发"(2021YFD1400100,2021YFD1400103)大连海关科研项目"检疫性病原菌白蜡鞘孢菌快速检测技术的研究与应用"(2024HK05)
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