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Detection of Salmonella Typhimurium on Spinach Using Phage-Based Magnetoelastic Biosensors

机译:基于噬菌体的磁弹性生物传感器检测菠菜中的鼠伤寒沙门氏菌

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Phage-based magnetoelastic (ME) biosensors have been studied as an in-situ, real-time, wireless, direct detection method of foodborne pathogens in recent years. This paper investigates an ME biosensor method for the detection of Salmonella Typhimurium on fresh spinach leaves. A procedure to obtain a concentrated suspension of Salmonella from contaminated spinach leaves is described that is based on methods outlined in the U.S. FDA Bacteriological Analytical Manual for the detection of Salmonella on leafy green vegetables. The effects of an alternative pre-enrichment broth (LB broth vs. lactose broth), incubation time on the detection performance and negative control were investigated. In addition, different blocking agents (BSA, Casein, and Superblock) were evaluated to minimize the effect of nonspecific binding. None of the blocking agents was found to be superior to the others, or even better than none. Unblocked ME biosensors were placed directly in a concentrated suspension and allowed to bind with Salmonella cells for 30 min before measuring the resonant frequency using a surface-scanning coil detector. It was found that 7 h incubation at 37 °C in LB broth was necessary to detect an initial spike of 100 cfu/25 g S. Typhimurium on spinach leaves with a confidence level of difference greater than 95% ( p < 0.05). Thus, the ME biosensor method, on both partly and fully detection, was demonstrated to be a robust and competitive method for foodborne pathogens on fresh products.
机译:近年来,已经研究了基于噬菌体的磁弹性(ME)生物传感器作为食源性病原体的实时,无线,直接检测方法。本文研究了一种ME生物传感器方法,用于检测新鲜菠菜叶上的鼠伤寒沙门氏菌。描述了一种从受污染的菠菜叶中获得沙门氏菌浓缩悬浮液的方法,该方法基于美国FDA细菌学分析手册中概述的用于在多叶绿色蔬菜上检测沙门氏菌的方法。研究了另一种预富集肉汤(LB肉汤与乳糖肉汤),孵育时间对检测性能和阴性对照的影响。另外,评估了不同的阻断剂(BSA,酪蛋白和超级嵌段)以最大程度地减少非特异性结合的影响。发现没有一种封闭剂优于其他封闭剂,甚至比没有更好。将未受阻的ME生物传感器直接置于浓缩悬浮液中,并与沙门氏菌细胞结合30分钟,然后使用表面扫描线圈检测器测量共振频率。发现在LB肉汤中于37°C孵育7 h对于检测100 cfu / 25 g S. Typhimurium在菠菜叶上的初始峰值很有必要,其置信水平的差异大于95%(p <0.05)。因此,ME生物传感器方法无论是部分检测还是全部检测,都被证明是一种针对新鲜产品上食源性病原体的稳健而竞争性方法。

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