一种快速检测含tst基因的金黄色葡萄球菌的纳米生物传感器

发布时间：2018-03-22 04:18

本文选题：单壁碳纳米管　切入点：荧光标记的核酸探针　出处：《生物技术通报》2017年12期 　论文类型：期刊论文

【摘要】：基于荧光能量共振转移(Fluorescence energy resonance transfer,FRET)原理利用单壁碳纳米管(Single-walled carbon nanotubes,SWCNTs)构建一种操作简单、成本低、灵敏度高和选择性强的纳米生物传感器快速检测含有tst基因金黄色葡萄球菌。将tst基因互补序列设计为探针,利用单壁碳纳米管联合羧基荧光素(FAM)标记的DNA分子探针(FAM-P)构建出FAM-P/SWCNTs复合检测体系。考察该体系对靶标序列和靶标菌的灵敏度和特异性。结果表明,该生物传感器对靶标序列的检测下限低至10 nmol/L,并在10-100 nmol/L范围内具有较好的线性关系(R2=0.994),对靶标菌的检测下限低至102,且在102-107CFU/m L范围内均呈现良好的线性关系(R2=0.999)。此外,在错配实验和对非靶标菌检测时,该传感器呈现了较高的特异性。本研究构建的FAM-P/SWCNTs体系能快速检测含tst基因金黄色葡萄球菌,并具有操作简单、灵敏度高、特异性强等优点,为食品检测、临床诊断等领域开辟了新途径。
[Abstract]:Based on the principle of fluorescence energy resonance transfer (fret), a novel single-walled carbon nanotube SWCNTs was constructed with simple operation and low cost. A sensitive and selective nanosensor for rapid detection of staphylococcus aureus containing tst gene was developed. The complementary sequence of tst gene was designed as a probe. The FAM-P/SWCNTs complex detection system was constructed by using single-walled carbon nanotubes (SWNTs) and carboxyl fluorescein (FAM) labeled DNA probe FAM-P.The sensitivity and specificity of the system to the target sequence and the target bacteria were investigated. The detection limit of the target sequence by the biosensor is as low as 10 nmol / L, and the linear relationship is good in the range of 10-100 nmol/L. The detection limit for the target bacteria is as low as 102, and the linear relationship is good in the range of 102-107CFU/m L. The FAM-P/SWCNTs system constructed in this study can detect staphylococcus aureus containing tst gene quickly and has the advantages of simple operation, high sensitivity, strong specificity and so on. For food detection, clinical diagnosis and other fields opened up a new way.
【作者单位】：中国医学科学院北京协和医学院医学生物学研究所中心实验室;长沙医学院基础医学院应用生物学教研室;
【基金】：高等学校博士学科点专项科研基金资助项目(20111106120056)
【分类号】：R446.5

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