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基于微流控的生物传感器快速检测食品中的沙门氏菌。

A microfluidic based biosensor for rapid detection of Salmonella in food products.

机构信息

University of Missouri-Columbia, Electrical and Computer Engineering, Columbia, MO, United States of America.

University of Missouri-Columbia, School of Veterinary Medicine, Columbia, MO, United States of America.

出版信息

PLoS One. 2019 May 14;14(5):e0216873. doi: 10.1371/journal.pone.0216873. eCollection 2019.

Abstract

An impedance based microfluidic biosensor for simultaneous and rapid detection of Salmonella serotypes B and D in ready-to-eat (RTE) Turkey matrix has been presented. Detection of Salmonella at a concentration as low as 300 cells/ml with a total detection time of 1 hour has been achieved. The sensor has two sensing regions, with each formed from one interdigitated electrode array (IDE array) consisting of 50 finger pairs. First, Salmonella antibody type B and D were prepared and delivered to the sensor to functionalize each sensing region without causing any cross contamination. Then the RTE Turkey samples spiked with Salmonella types B and D were introduced into the biosensor via the antigen inlet. The response signal resulted from the binding between Salmonella and its specific antibody demonstrated the sensor's ability to detect a single type of pathogen, and multiple pathogens simultaneously. In addition, the biosensor's selectivity was tested using non-specific binding of E. coli O157 and E. coli DH5 Alpha while the IDE array was coated with the Salmonella antibody. The results also showed the sensor is capable to differentiate low concentration of live Salmonella cells from high concentration of dead Salmonella cells, and high concentration of E. coli cells. A detailed study on antibody immobilization that includes antibody concentration, antibody coating time (0.5-3 hours) and use of cross-linker has been performed. The study showed that Salmonella antibody to Salmonella antigen is not a factor of antibody concentration after electrodes were saturated with antibody, while the optimal coating time was found to be 1.5 hours, and the use of cross-linker has improved the signal response by 45-60%.

摘要

已经提出了一种基于阻抗的微流控生物传感器,用于同时快速检测即食(RTE)火鸡基质中的 B 型和 D 型沙门氏菌。已经实现了以低至 300 个细胞/ml 的浓度检测沙门氏菌,总检测时间为 1 小时。该传感器有两个传感区域,每个区域都由一个由 50 对指状对组成的叉指电极阵列(IDE 阵列)形成。首先,制备了 B 型和 D 型沙门氏菌抗体,并将其递送到传感器上,以在不引起任何交叉污染的情况下使每个传感区域功能化。然后,将含有 B 型和 D 型沙门氏菌的 RTE 火鸡样品通过抗原入口引入生物传感器。传感器的响应信号来自沙门氏菌与其特异性抗体之间的结合,证明了传感器能够检测单一类型的病原体和多种病原体。此外,通过在 IDE 阵列上涂覆沙门氏菌抗体,测试了生物传感器的选择性,以测试非特异性结合的大肠杆菌 O157 和大肠杆菌 DH5 Alpha。结果还表明,该传感器能够区分低浓度的活沙门氏菌细胞与高浓度的死沙门氏菌细胞,以及高浓度的大肠杆菌细胞。还对包括抗体浓度、抗体涂层时间(0.5-3 小时)和使用交联剂在内的抗体固定化进行了详细研究。研究表明,在电极被抗体饱和后,沙门氏菌抗体与沙门氏菌抗原之间的关系不是抗体浓度的因素,而最佳涂层时间被发现为 1.5 小时,使用交联剂可使信号响应提高 45-60%。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/65fb/6516674/c2de79e17daf/pone.0216873.g001.jpg

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