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基于碳纳米管/金纳米粒子纳米复合材料的扫描磁免疫分析用于检测鼠伤寒沙门氏菌血清型。

Scano-magneto immunoassay based on carbon nanotubes/gold nanoparticles nanocomposite for Salmonella enterica serovar Typhimurium detection.

机构信息

School of Bioresources and Technology, King Mongkut's University of Technology Thonburi, Bangkhuntien, Bangkok 10150, Thailand.

出版信息

Biosens Bioelectron. 2012 Oct-Dec;38(1):157-62. doi: 10.1016/j.bios.2012.05.018. Epub 2012 May 29.

Abstract

To improve sensitivity of S. enterica serovar Typhimurium detection, multiwalled carbon nanotubes (MWCNTs) and gold nanoparticles (AuNPs) were combined and used as a label to amplify signal in a scanometric based assay. In this study, the MWCNTs/AuNPs nanocomposite was fabricated by directly assemble of Au(3+) to MWCNTs and allowed growing of AuNPs along the MWCNTs surface. This MWCNTs/AuNPs nanocomposite was then attached to anti-S. typhimurium antibody (MWCNTs/AuNPs/Ab(1)) and used as a detecting molecule. Upon binding to Salmonella, they were pre-concentrated by magenetic beads/antibody (MBs/Ab(2)) forming a sandwich immuno-complex which is later spotted on a nitrocellulose membrane coated slide. Silver reduction was applied to amplify signal. The detection limit of 42CFU/ml was achieved when 2% BSA was used as a blocking agent. Given different types of real samples testing, chicken broth was found to give lowest detection limit, followed by orange juice low fat and whole milk. Selectivity testing was performed by using Escherichia coli as interference and found slightly cross-reactivity which could be due to specificity of the Ab used. By virtue of using a slide for multi-samples spotting and a flatbed scanner for signal-read out acquisition, this scano-magneto immunoassay could enable low-cost detection as well as high throughput screening.

摘要

为了提高肠炎沙门氏菌检测的灵敏度,将多壁碳纳米管(MWCNTs)和金纳米颗粒(AuNPs)结合起来,作为一种标记物,在基于扫描计的测定中放大信号。在这项研究中,MWCNTs/AuNPs 纳米复合材料是通过直接将 Au(3+)组装到 MWCNTs 上,并允许 AuNPs 在 MWCNTs 表面生长而制成的。然后,将这种 MWCNTs/AuNPs 纳米复合材料附着到抗鼠伤寒沙门氏菌抗体(MWCNTs/AuNPs/Ab(1))上,用作检测分子。与沙门氏菌结合后,它们被磁珠/抗体(MBs/Ab(2))预先浓缩,形成三明治免疫复合物,然后将其点在涂有硝酸纤维素膜的载玻片上。应用银还原法放大信号。当使用 2%BSA 作为封闭剂时,检测限达到 42CFU/ml。在对不同类型的实际样品进行测试时,发现鸡肉汤的检测限最低,其次是低脂肪橙汁和全脂牛奶。通过使用大肠杆菌作为干扰物进行选择性测试,发现了轻微的交叉反应性,这可能是由于所用抗体的特异性。通过在载玻片上进行多样本点样和平板扫描仪进行信号读取采集,这种扫描磁免疫测定法既可以实现低成本检测,又可以进行高通量筛选。

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