摘要
建立一种镰刀菌核酸检测方法,为镰刀菌田间复合侵染早期诊断提供支持。本研究设计并合成18S-1、28S-1共2组镰刀菌属特异性引物,选取层出镰刀菌(1-CC)、腐皮镰刀菌(15-FP)和禾谷镰刀菌(36-HG)等10株供试菌株开展引物筛选试验;筛选获得最佳引物组合后,搭配其他配套试剂构建针对镰刀菌属的通用核酸快速检测等温扩增荧光体系。采用该检测体系测定镰刀菌核酸的检出限,并以常规PCR技术作为参照方法,开展灵敏度对比评价。结果表明,18S-1引物组扩增效率高、时间短,宜作为最佳引物组应用到后续试验中;等温扩增荧光法对镰刀菌有特异性扩增,检测限为10^(-5)浓度,与常规PCR方法(10^(-3)浓度)相比灵敏度较高。综上,该方法具有良好的特异性和较高的灵敏度,能够用于镰刀菌的快速检测。
To establish nucleic acid detection method for Fusarium,providing support for the early diagnosis of Fusarium field complex infection.2 groups of Fusarium genus specific primers,namely 18S-1 and 28S-1,were designed and synthesized.10 test strains such as Fusarium proliferatum(1-CC),Fusarium solani(15-FP),and Fusarium graminearum(36-HG)were selected for primer screening experiments.After screening for the best primer combination,a universal nucleic acid rapid detection isothermal amplification fluorescence system for Fusarium was constructed in combination with other supporting reagents.The detection limit of Fusarium nucleic acid was determined by using this detection system,and the sensitivity comparison and evaluation were carried out with conventional PCR technology as the reference method.The results showed that the 18S-1 primer group had high amplification efficiency and short time,and was suitable to be used as the best primer group for subsequent experiments.Isothermal amplification fluorescence method had specific amplification for Fusarium,with a detection limit of 10^(-5)concentration,and higher sensitivity compared with the common PCR method(10^(-3)concentration).In conclusion,this method has good specificity and high sensitivity,and can be used for the rapid detection of Fusarium.
作者
梁振瑞
姜仁普
张薇
李成云
许绍坤
段真文
Liang Zhenrui;Jiang Renpu;Zhang Wei;Li Chengyun;Xu Shaokun;Duan Zhenwen(Yunnan Rare Biotechnology Co.,Ltd.,Kunming 650000,China)
出处
《安徽农学通报》
2026年第3期96-98,共3页
Anhui Agricultural Science Bulletin
基金
云南省科技厅项目“分子检测诊断产品关键技术研发”(202102AA310028)。
关键词
镰刀菌
灵敏度
特异性
等温扩增荧光法
Fusarium
sensitivity
specificity
isothermal amplification fluorescence method
