基于DNAzyme的荧光生物传感器快速检测金黄色葡萄球菌研究

木卡地斯·米吉提; 王辉; 潘东霞; 申瑶; 杨亮

doi:10.7671/j.issn.1001-411X.202405008

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

摘要

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1~1×10⁷ 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。

Abstract:
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 1−1×10⁷ 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.

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