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用于检测实际样本中食源性致病菌的微纳生物声系统。

Micro-nano-bio acoustic system for the detection of foodborne pathogens in real samples.

机构信息

Institute of Molecular Biology and Biotechnology-FORTH, 100 N. Plastira Str., Heraklion 70013, Greece.

University of Pardubice, 573 Studentska, Pardubice 532 10, Czech Republic.

出版信息

Biosens Bioelectron. 2018 Jul 15;111:52-58. doi: 10.1016/j.bios.2018.03.056. Epub 2018 Mar 28.

Abstract

The fast and efficient detection of foodborne pathogens is a societal priority, given the large number of food-poisoning outbreaks, and a scientific and technological challenge, given the need to detect as little as 1 viable cell in 25 gr of food. Here, we present the first approach that achieves the above goal, thanks to the use of a micro/nano-technology and the detection capability of acoustic wave sensors. Starting from 1 Salmonella cell in 25 ml of milk, we employ immuno-magnetic beads to capture cells after only 3 h of pre-enrichment and subsequently demonstrate efficient DNA amplification using the Loop Mediated Isothermal Amplification method (LAMP) and acoustic detection in an integrated platform, within an additional ½ h. The demonstrated 4 h sample-to-analysis time comes as a huge improvement to the current need of few days to obtain the same result. In addition, the work presents the first reported Lab-on-Chip platform that comprises an acoustic device as the sensing element, exhibiting impressive analytical features, namely, an acoustic limit of detection of 2 cells/μl or 3 aM of the DNA target and ability to detect in a label-free manner dsDNA amplicons in impure samples. The use of food samples together with the incorporation of the necessary pre-enrichment step and ability for multiple analysis with an internal control, make the proposed methodology highly relevant to real-world applications. Moreover, the work suggests that acoustic wave devices can be used as an attractive alternative to electrochemical sensors in integrated platforms for applications in food safety and the point-of-care diagnostics.

摘要

鉴于食源性疾病暴发的数量众多,快速、高效地检测食源性病原体已成为当务之急,这是一个科学技术方面的挑战,因为需要在 25 克食物中检测到少至 1 个活细胞。在这里,我们首次提出了一种基于微纳技术和声波传感器检测能力的方法来实现这一目标。从 25 毫升牛奶中的 1 个沙门氏菌细胞开始,我们仅在预富集 3 小时后使用免疫磁珠捕获细胞,随后使用环介导等温扩增(LAMP)方法进行有效的 DNA 扩增,并在集成平台中进行声学检测,整个过程在另外的半小时内完成。与目前获得相同结果需要数天的时间相比,所展示的 4 小时样本分析时间有了很大的改进。此外,这项工作还首次报道了一种包含声学器件作为传感元件的芯片实验室平台,该平台具有令人印象深刻的分析特性,即声探测极限为 2 个细胞/μl 或 3 aM 的 DNA 靶标,并且能够以无标记的方式检测不纯样品中的 dsDNA 扩增子。该方法使用了食品样本,并结合了必要的预富集步骤以及使用内部控制进行多次分析的能力,使其非常适用于实际应用。此外,这项工作表明,声波器件可作为集成平台中电化学传感器的一种有吸引力的替代品,用于食品安全和即时诊断应用。

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