Optical Detection of Listeria in the Chilled Food Environment using Bionanosensors

使用生物纳米传感器光学检测冷冻食品环境中的李斯特菌

• 批准号: BB/R00899X/1
• 负责人: Karen Faulds
• 金额: $ 46.58万
• 依托单位: University of Strathclyde
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2018
• 资助国家: 英国
• 起止时间: 2018 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FR00899X%2F1
• 关键词: Optical; Detection; Listeria; Chilled; Food

This programme of research involves the development of an innovative bionanosensor with superior performance for the detection of bacterial pathogens in a sensitive, quantitative and multiplexed manner for use in chilled food production. This will involve developing nanoparticle based analytical technology for the detection of multiple bacterial pathogens (L. monocytogenesis and enterobacteriaceae) associated with food contamination in the chilled food industry. Current methods for detecting bacteria are time consuming (1-2 days in the case of bacteria culturing on selective media, but in reality 7 day turnaround for food production facilities where laboratories are located offsite), expensive and require specialised personnel and equipment. Therefore, there is a strong need for faster, simpler and reliable isolation and detection of bacterial pathogens that can be carried out in the production environment. To address this need we will develop a simple, portable detection platform capable of point of use (POU) detection for bacteria.Successful bacteria detection in food production is crucial for the health of the general public. Within the chilled food sector, Listeria is of particular concern since it is an unusually resilient bacterium which can survive in temperatures from 4 to 42 degrees C. Listeria monocytogenes causes one of the most severe forms of foodborne infection with an overall mortality rate of 30 %, rising to 40 % in vulnerable individuals. Those most at risk of acquiring the disease are pregnant women, the elderly and the immuno-compromised and in some cases listeriosis can develop into meningitis and septicaemia if infection of the blood occurs. However, outbreaks of Listeria are rare due to strict controls on food safety however our industrial partner, Bradgate Bakery, who manufacture sandwiches and salads, require the ability to locate, detect and contain any contamination within food production lines.The research involves the use of an optical detection technique called Raman scattering which will be developed for the POU detection of bacterial pathogens. If light of a particular wavelength is directed onto a molecule then some of the scattered light will change wavelength. This change in wavelength is related to the structure of the molecules and provides a molecular fingerprint that can be used for definitive identification. However Raman scattering is an intrinsically weak process and the signal can be greatly enhanced if the molecule is coloured and is adsorbed onto a roughened metal surface (surface enhanced resonance Raman). The metal can be thought of as essentially amplifying the Raman scattering from a molecule on the surface and in this case will take the form of metal nanoparticles. Since a fingerprint unique to the molecule is produced, the composition of mixtures can easily be identified without separation. A novel bionanosensor for the detection of multiple bacterial pathogens, namely L. monocytogenesis and enterobacteriaceae in one assay combined with enhanced Raman detection will be developed. This will use magnetic nanoparticles which have a biomolecule on the surface known as a lectin which will bind to the surface of bacteria. This will allow isolation and separation of bacteria from the surrounding medium upon application of a magnetic. Additionally, silver nanoparticles which are functionalised with a coloured molecule or label, resulting in intense surface enhanced Raman signals, and a biomolecule which will bind specifically to a particular strain of bacteria (antibody or aptamer) will be added. When the correct bacteria are present binding will occur resulting in magnetic isolation and concentration of the bacteria from the matrix. By using a different label for each bacteria, a unique spectrum will be achieved for each bacteria allowing multiple to be detected simultaneously. A portable Raman spectrometer will then be used to detect the bacteria present.

这项研究计划涉及开发一种创新的生物纳米传感器，具有上级性能，可用于检测冷冻食品生产中使用的细菌病原体，其检测方式灵敏，定量和多重。这将涉及开发基于纳米颗粒的分析技术，用于检测多种细菌病原体（L。单核细胞生成和肠杆菌科）与冷冻食品行业的食品污染有关。目前用于检测细菌的方法是耗时的（在选择性培养基上培养细菌的情况下为1-2天，但实际上对于实验室位于场外的食品生产设施而言为7天的周转时间），昂贵并且需要专业人员和设备。因此，迫切需要可以在生产环境中进行的细菌病原体的更快、更简单和可靠的分离和检测。为了满足这一需求，我们将开发一种简单、便携的检测平台，能够对细菌进行使用点（POU）检测。在食品生产中成功进行细菌检测对公众健康至关重要。在冷冻食品行业，李斯特菌特别令人关注，因为它是一种异常有弹性的细菌，可以在4至42摄氏度的温度下生存。单核细胞增生李斯特菌导致最严重形式的食源性感染之一，总体死亡率为30%，在脆弱个体中上升至40%。孕妇、老人和免疫力低下的人最容易患上这种疾病，在某些情况下，如果血液受到感染，脑膜炎和败血症会发展成脑膜炎和败血症。然而，由于严格控制食品安全，李斯特菌的爆发是罕见的，但我们的工业合作伙伴，生产三明治和沙拉的Bradgate Bakery，需要能够定位，检测和控制食品生产线内的任何污染。这项研究涉及使用一种称为拉曼散射的光学检测技术，该技术将用于细菌病原体的POU检测。如果特定波长的光被引导到分子上，那么一些散射光将改变波长。这种波长的变化与分子的结构有关，并提供了可用于最终鉴定的分子指纹。然而，拉曼散射本质上是弱过程，并且如果分子被着色并被吸附到粗糙的金属表面上，则信号可以大大增强（表面增强共振拉曼）。金属可以被认为是基本上放大来自表面上的分子的拉曼散射，并且在这种情况下将采取金属纳米颗粒的形式。由于产生了分子特有的指纹，混合物的组成可以很容易地识别而无需分离。建立了一种检测多种细菌病原体的新型生物纳米传感器。将开发与增强型拉曼检测相结合的一种测定中的单核细胞生成和肠杆菌科的方法。这将使用磁性纳米颗粒，其表面上具有被称为凝集素的生物分子，其将结合到细菌表面。这将允许在施加磁性时从周围介质中分离和分离细菌。此外，将添加用有色分子或标记官能化的银纳米颗粒，导致强烈的表面增强拉曼信号，以及将特异性结合特定细菌菌株的生物分子（抗体或适体）。当存在正确的细菌时，将发生结合，导致细菌从基质中的磁性分离和浓缩。通过对每种细菌使用不同的标记，将为每种细菌实现独特的光谱，从而允许同时检测多种细菌。然后将使用便携式拉曼光谱仪检测存在的细菌。

项目成果

Surface enhanced Raman scattering for the multiplexed detection of pathogenic microorganisms: towards point-of-use applications.

• DOI: 10.1039/d1an00865j
• 发表时间: 2021-10-11
• 期刊: The Analyst
• 作者: Berry ME;Kearns H;Graham D;Faulds K
• 通讯作者: Faulds K

Tomographic Imaging and Localization of Nanoparticles in Tissue Using Surface-Enhanced Spatially Offset Raman Spectroscopy.

• DOI: 10.1021/acsami.2c05611
• 发表时间: 2022-07-20
• 期刊: ACS APPLIED MATERIALS & INTERFACES
• 影响因子: 9.5
• 作者: Berry, Matthew E.;McCabe, Samantha M.;Sloan-Dennison, Sian;Laing, Stacey;Shand, Neil C.;Graham, Duncan;Faulds, Karen
• 通讯作者: Faulds, Karen

Evaluating nanoparticle localisation in glioblastoma multicellular tumour spheroids by surface enhanced Raman scattering.

• DOI: 10.1039/d3an00751k
• 发表时间: 2023-07-10
• 期刊: The Analyst

From Raman to SESORRS: moving deeper into cancer detection and treatment monitoring.

• DOI: 10.1039/d1cc04805h
• 发表时间: 2021-11-23
• 期刊: Chemical communications (Cambridge, England)
• 作者: Sloan-Dennison S;Laing S;Graham D;Faulds K
• 通讯作者: Faulds K

Surface Design for Immobilization of an Antimicrobial Peptide Mimic for Efficient Anti-Biofouling

• DOI: 10.1002/chem.202000746
• 发表时间: 2020-04-21
• 期刊: CHEMISTRY-A EUROPEAN JOURNAL
• 影响因子: 4.3
• 作者: Hasan, Abshar;Lee, Kyueui;Lau, King Hang Aaron
• 通讯作者: Lau, King Hang Aaron