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基于表面等离子体共振成像的食源性沙门氏菌无标记筛选

Label-free screening of foodborne Salmonella using surface plasmon resonance imaging.

机构信息

United States Department of Agriculture, Agricultural Research Service, U.S. National Poultry Research Center, 950 College Station Rd., Athens, GA, 30605, USA.

出版信息

Anal Bioanal Chem. 2018 Sep;410(22):5455-5464. doi: 10.1007/s00216-017-0810-z. Epub 2017 Dec 26.

Abstract

It is estimated that 95% of the foodborne infections are caused by 15 major pathogens. Therefore, rapid and effective multiplex screening techniques for these pathogens with improved efficiencies could benefit public health at lower costs. Surface plasmon resonance imaging (SPRi) provides a label-free, multiplex analytical platform for pathogen screening. In this study, we have developed a singleplex immunoassay for Salmonella to evaluate the potential of SPRi in pathogen detection. Anti-Salmonella and control ligands were arrayed onto the SPRi sensor chip in a microarray format. The influences of ligand immobilization pH and concentration were optimized, and a pause flow protocol was adopted to improve assay rapidity and sensitivity. The method shows good specificity against 6 non-Salmonella species and was able to detect 5 of 6 Salmonella serotypes, including 3 serotypes most frequently associated with outbreaks. Limits of detection were found to be 2.1 × 10 CFU/mL in phosphate-buffered saline and 7.6 × 10 CFU/mL in the presence of chicken rinse matrix with 8.9 × 10 CFU/mL of indigenous microflora. The condition of antibody array regeneration was optimized for sequential sample injections. Finally, the SPRi immunoassay was used to detect Salmonella directly from artificially spiked chicken carcass rinse samples. As low as 6.8 CFU/mL of Salmonella could be detected after overnight enrichment in buffered peptone water, demonstrating the potential in streamlined pathogen screening with minimal sample preparation and without detection labels. Graphical abstract ᅟ.

摘要

据估计,95%的食源性感染是由 15 种主要病原体引起的。因此,开发能够提高效率的这些病原体的快速有效的多重筛选技术,可以以更低的成本造福公共健康。表面等离子体共振成像(SPRi)为病原体筛选提供了一种无标记的多重分析平台。在这项研究中,我们开发了一种用于检测沙门氏菌的单重免疫分析方法,以评估 SPRi 在病原体检测中的潜力。将抗沙门氏菌和对照配体以微阵列的形式排列在 SPRi 传感器芯片上。优化了配体固定化 pH 值和浓度的影响,并采用暂停流动方案来提高分析的快速性和灵敏度。该方法对 6 种非沙门氏菌具有良好的特异性,能够检测到 6 种沙门氏菌血清型中的 5 种,包括与暴发最相关的 3 种血清型。在磷酸盐缓冲液中的检测限为 2.1×10 CFU/mL,在含有鸡冲洗基质的情况下为 7.6×10 CFU/mL,其中土著微生物群的检测限为 8.9×10 CFU/mL。优化了抗体阵列再生条件,以实现连续样品注射。最后,使用 SPRi 免疫分析直接从人工污染鸡胴洗液样品中检测沙门氏菌。在缓冲蛋白胨水中过夜富集后,可检测到低至 6.8 CFU/mL 的沙门氏菌,证明了在简化病原体筛选方面具有潜力,该方法需要的样品制备最少,且无需检测标签。

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