基于DNAzyme的荧光生物传感器快速检测金黄色葡萄球菌研究  

Research on rapid detection of Staphylococcus aureus by fluorescent biosensor based on DNAzyme

作　　者：木卡地斯·米吉提 王辉 潘东霞 申瑶 杨亮[1] MUKADDAS Mijit;WANG Hui;PAN Dongxia;SHEN Yao;YANG Liang(Institute of Animal Sciences,Chinese Academy of Agricultural Sciences/State Key Laboratory of Animal Nutrition and Feeding,Beijing 100193,China)

机构地区：[1]中国农业科学院北京畜牧兽医研究所/畜禽营养与饲养全国重点实验室,北京100193

出　　处：《华南农业大学学报》2024年第5期764-771,共8页Journal of South China Agricultural University

基　　金：广东省重点研发计划(2023B0202140001-3);新一代人工智能国家科技重大专项(2021ZD0113801);山东省重点研发计划(2022TZXD0016-1)。

摘　　要：【目的】为解决传统方法检测金黄色葡萄球菌耗时长、操作复杂等问题,研发一种基于脱氧核酶(DNAzyme)的荧光生物传感器,实现金黄色葡萄球菌的快速检测。【方法】将特异性DNAzyme和互补链Subatrate相结合制成荧光生物传感器,并对荧光生物传感器进行生物材料浓度和溶液pH优化,并对金黄色葡萄球菌、大肠埃希菌、芽孢杆菌、无乳链球菌、变形杆菌等进行特异性检测;最后对牛奶样品进行基于DNAzyme的荧光生物传感器的试验验证。【结果】基于DNAzyme的荧光生物传感器在pH为6.8时,3 min内可以实现对金黄色葡萄球菌的检测,线性范围为1~1×10^(7) cfu·mL^(−1),最低检测限为1 cfu·mL^(−1)。【结论】基于DNAzyme的荧光生物传感器解决了传统检测方法耗时长、操作复杂等问题,实现了对金黄色葡萄球菌的快速检测,具有重要的应用价值。【Objective】To solve the problems of time-consuming and complex operation in traditional methods of detecting Staphylococcus aureus,a fluorescent biosensor based on deoxyribonuclease(DNAzyme)was developed to achieve rapid detection of S.aureus.【Method】The fluorescence biosensor was made by combining DNAzyme with specific complementary chain substrate,and optimized for the concentration of biomaterials and pH of solution.Then,the specific detection of five different bacteria such as S.aureus,Escherichia coli,Bacillus,Streptococcus agalactiae and Proteus was performed.Finally,the DNAzyme-based fluorescent biosensor was tested and verified on milk samples.【Result】The fluorescent biosensor based on DNAzyme can detect S.aureus within 3 min at a pH of 6.8,with a linear range of 1−1×10^(7) cfu·mL^(−1) and a minimum detection limit of 1 cfu·mL^(−1).【Conclusion】The fluorescent biosensor based on DNAzyme solves the problems of time-consuming and complex operation of traditional detection methods,and realizes the rapid detection of S.aureus in milk,which has significant practical value.

关 键 词：金黄色葡萄球菌 DNAZYME 分子信标 荧光生物传感器 

分 类 号：S815[农业科学—畜牧学]