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基于金和铁纳米粒子的生物条形码技术检测金黄色葡萄球菌蛋白 A

Bio-barcode technology for detection of Staphylococcus aureus protein A based on gold and iron nanoparticles.

机构信息

Nano Biotechnology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran.

Nano Biotechnology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran.

出版信息

Int J Biol Macromol. 2019 Mar 1;124:1256-1263. doi: 10.1016/j.ijbiomac.2018.11.123. Epub 2018 Nov 14.

Abstract

S. aureus is one of important causes of disease, food poisoning in humans and animals. The generally methods for detection of S. aureus is time consuming. Therefore, a new method is necessary for rapid, sensitive and specific diagnosis of S. aureus. In the present study, two probes and a Bio-barcode DNA were designed for detection of S. aureus (Protein A). Firstly, magnetic nanoparticle (MNPs) and gold nanoparticle (AuNPs) were synthesized at 80 °C and 100 °C, respectively. The AuNPs and the MNPs were functionalized with probe1, Bio-barcode DNA and probe2, respectively. Target DNA was added into the nanomaterial's system containing bio-barcode DNA-AuNPs-probe1 and probe2-MNPs to formed bio-barcode DNA-AuNPs-probe1-target DNA-probe2-MNPs complex. The bio-barcode DNA-AuNPs-probe1-target DNA-probe2-MNPs complex was separated with magnetic field. Finally, the bio-barcode DNA was released from surface of complex using DTT (0.8 M) and there was isolated of nanoparticles by magnetic field and centrifuge. The fluorescence intensity of bio-barcode DNA was measured in different concentrations of S. aureus (10 to 10 CFU mL) by fluorescence spectrophotometry. The results showed that standard curve was linearly from 10 to 10 CFU mL. Limit of detection of bio-barcode assay for both PBS and real samples was 86 CFU mL.

摘要

金黄色葡萄球菌是人类和动物疾病和食物中毒的重要原因之一。目前用于检测金黄色葡萄球菌的方法通常比较耗时。因此,需要一种新的方法来快速、灵敏和特异性地诊断金黄色葡萄球菌。本研究设计了两种探针和一个生物条码 DNA 来检测金黄色葡萄球菌(蛋白 A)。首先,在 80°C 和 100°C 下分别合成了磁性纳米颗粒(MNPs)和金纳米颗粒(AuNPs)。AuNPs 和 MNPs 分别用探针 1、生物条码 DNA 和探针 2 进行功能化。将靶 DNA 加入含有生物条码 DNA-AuNPs-探针 1 和探针 2-MNPs 的纳米材料体系中,形成生物条码 DNA-AuNPs-探针 1-靶 DNA-探针 2-MNPs 复合物。利用磁场将生物条码 DNA-AuNPs-探针 1-靶 DNA-探针 2-MNPs 复合物分离。最后,用 DTT(0.8 M)从复合物表面释放出生物条码 DNA,并利用磁场和离心将纳米粒子分离。通过荧光分光光度法测量不同浓度金黄色葡萄球菌(10 到 10⁶ CFU/mL)的生物条码 DNA 的荧光强度。结果表明,标准曲线在 10 到 10⁶ CFU/mL 之间呈线性关系。生物条码分析在 PBS 和实际样品中的检测限均为 86 CFU/mL。

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