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基于过滤的 SERS 映射平台的开发,用于特异性筛选肠炎沙门氏菌血清型肠炎亚种。

Development of a filtration-based SERS mapping platform for specific screening of Salmonella enterica serovar Enteritidis.

机构信息

Department of Food Science, University of Massachusetts Amherst, Amherst, MA, 01002, USA.

出版信息

Anal Bioanal Chem. 2019 Nov;411(29):7899-7906. doi: 10.1007/s00216-019-02204-3. Epub 2019 Nov 20.

Abstract

The presence of Salmonella in natural freshwater and drinking water is a leading cause of intestinal illness all over the world; thus, the detection of Salmonella in water is of great importance to public health. The objective of this study is to develop a rapid screening method for the detection of Salmonella enterica serovar Enteritidis in water involving surface-enhanced Raman spectroscopy (SERS), aptamers, and filtration. SERS offers a great alternative to traditional methods of pathogen detection, with a simplified detection assay and shortened detection time. The specific capturing and labeling of Salmonella Enteritidis are realized by a specific single-stranded DNA aptamer, which is modified with an additional chain of adenine and fluorescein (FAM) and used as presence/absence indicator of Salmonella Enteritidis. By incorporating a vacuum filtration system, bacterial cells recognized by the specific aptamer are concentrated onto a membrane. With additional filtration of gold nanoparticles, the aptamer signals were captured and used to construct a SERS mapping indicating the presence and absence of target bacterial strains with potential quantitative capability. The specificity of the method was validated by using other strains of bacteria such as Escherichia coli and Listeria monocytogenes. The sensitivity of the method goes down to 10 CFU/mL for 1 mL of sample with a total detection and analyzing time within 3 h. This study demonstrates the capability of the filtration-based SERS platform for detecting Salmonella Enteritidis in various aqueous matrices such as distilled water and rinsing water from fresh produce with high selectivity and sensitivity. Graphical abstract.

摘要

在天然淡水和饮用水中存在沙门氏菌是全世界肠道疾病的主要原因;因此,水中沙门氏菌的检测对公众健康非常重要。本研究的目的是开发一种涉及表面增强拉曼光谱(SERS)、适体和过滤的快速筛选方法,用于检测水中的肠炎沙门氏菌血清型肠炎亚种。SERS 为病原体检测的传统方法提供了很好的替代方法,具有简化的检测分析和缩短的检测时间。通过用额外的腺嘌呤和荧光素(FAM)链修饰特异性单链 DNA 适体,实现了肠炎沙门氏菌的特异性捕获和标记,该适体被用作肠炎沙门氏菌的存在/不存在指示物。通过加入真空过滤系统,被特异性适体识别的细菌细胞被浓缩到膜上。通过进一步过滤金纳米粒子,捕获适体信号并用于构建 SERS 图谱,该图谱指示具有潜在定量能力的目标细菌菌株的存在和不存在。通过使用其他细菌菌株,如大肠杆菌和单核细胞增生李斯特菌,验证了该方法的特异性。该方法的灵敏度可低至 10 CFU/mL(1 mL 样品),总检测和分析时间在 3 小时内。本研究证明了基于过滤的 SERS 平台在检测各种水基基质(如蒸馏水和新鲜农产品的冲洗水)中的肠炎沙门氏菌的能力,具有高选择性和灵敏度。

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