交链孢酚适配体荧光试纸条检测方法的建立及应用  

Establishment and Application of Aptamer-based Fluorescent Test Strip Method for the Detection of Alternariol

作　　者：方梦旗 赵颖 王子晨 贾文昊 王辉[2] 栾云霞[1] FANG Mengqi;ZHAO Ying;WANG Zichen;JIA Wenhao;WANG Hui;LUAN Yunxia(Agricultural Product Quality and Safety Risk Assessment Laboratory(Beijing)of the Ministry of Agriculture and Rural Affairs,Institute of Quality Standard and Testing Technology,Beijing Academy of Agriculture and Forestry Sciences,Beijing 100097,China;Biomedical Department of Beijing City University,Beijing 100191,China)

机构地区：[1]北京市农林科学院质量标准与检测技术研究所,农业农村部农产品质量安全风险评估实验室(北京),北京100097 [2]北京城市学院生物医药学部,北京100191

出　　处：《生物技术进展》2024年第6期1024-1031,共8页Current Biotechnology

基　　金：北京市农林科学院科技创新专项建设资助项目(KJCX20230437,KJCX20220410,KJCX20230309)。

摘　　要：交链孢酚(alternariol,AOH),作为交链孢霉菌产生的一种有毒代谢产物,通过侵染小麦、番茄、樱桃和柑橘等农产品,不仅造成经济损失,还对人类健康构成威胁。基于竞争原理,构建了一种针对AOH的灵敏且特异性强的核酸适配体荧光试纸条检测体系。对适配体浓度、pH和缓冲液等条件进行优化后,进一步评价其准确性和特异性等性能,并应用于小麦样品中AOH的含量测定。在最优条件下,其检测限可达1μg·L^(-1),检测范围为1~1000μg·L^(-1),线性相关系数R^(2)=0.9792,仅需20 min即可完成检测。在实际样品分析中的回收率为96.6%~114.3%,相对标准偏差为2.6%~9.0%。利用核酸适配体作为识别元件,开发了一种简便、快速、准确的适合现场快速定量检测AOH的新方法,为其他真菌毒素快速检测技术的研发提供了参考。Alternariol(AOH)is a toxic metabolite produced by the fungus Alternariol.By infecting wheat,tomato,cherry,citrus and other agricultural products,Streptomyces alternans not only causes economic losses but also endangers human health.In this study,based on the principle of competition,a sensitive and specific aptamer fluorescence test strip for AOH detection was constructed.After optimizing the aptamer concentration,pH value and buffer,the accuracy and specificity of the aptamer were further evaluated and applied to the determination of AOH in wheat samples.Under the optimal conditions,the detection limit was 1μg·L^(-1),the detection range was 1~1000μg·L^(-1),the linear correlation coefficient R^(2)=0.9792,the detection could be completed in only 20 minutes,the recovery rate in the actual sample analysis was 96.6%~114.3%,the relative standard deviation was 2.6%~9.0%.In this study,a simple,rapid and accurate new method for rapid quantitative detection of AOH was developed using aptamer as a recognition element,which would provide a reference for the development of other mycotoxin rapid detection technologies.

关 键 词：核酸适配体 荧光侧流层析试纸条 交链孢酚 小麦 

分 类 号：Q524[生物学—生物化学] TS207.53[轻工技术与工程—食品科学]