A fully integrated microdevice for capturing, amplification, and colorimetric detection of foodborne pathogens
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TECHNICAL PAPER
A fully integrated microdevice for capturing, amplification, and colorimetric detection of foodborne pathogens Hoang Chau La1 • Nae Yoon Lee1 Received: 26 February 2020 / Accepted: 8 May 2020 Ó Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract In the present study, we integrated laboratory functionalities of sample preparation, nucleic acid amplification, and colorimetric detection into a microdevice for the facile foodborne pathogen detection from bulk liquid samples. The developed microdevice consists of micropillars engraved in a microchamber. In order to capture pathogens from the high throughput sample, boronic acid was decorated on the surface of micropillars to exploit the high affinity of this functional group toward cis-diols of moieties constructing the bacterial outer membrane. Using disposable syringes, the surface modification process for the dense distribution of the boronic acid group was readily achieved at room temperature within 3 h. The success of surface modification was verified by the color changes and the fluorescence signals of the remnant alizarin red sodium. Escherichia coli O157:H7 involving green fluorescence protein (E. coli O157:H7 GFP) was used to evaluate the capture capability of the device at three inoculum bacterial concentrations. The temperature in the reaction zone was carefully investigated to ensure appropriate thermal conditions for performing chamber-type polymerase chain reaction. According to the results, the 584-bp sequence of the Shiga toxin gene of the pathogen was successfully amplified within 2 h. The amplicon was then visualized by the brown color of NaBH4-induced silver nanoparticles. Based on the results, it can be concluded that the proposed microdevice is a promising platform for robust detection of the foodborne pathogens in bulk liquid samples and is applicable in food industry, public healthcare, and environmental monitoring.
1 Introduction Rapid identification of pathogenic microorganisms is vital for monitoring an epidemic at its early stage of development. However, conventional methods for pathogen identification are time-consuming and have low sensitivity (Mairhofer et al. 2009). The recently developed nucleic acid-based, biosensor-based, and immunological-based approaches have successfully overcome these limitations (Law et al. 2014). Yet, these novel approaches require robust and sophisticated laboratory facilities and welltrained technicians. Moreover, the transfer of the bacterial
Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00542-020-04877-8) contains supplementary material, which is available to authorized users. & Nae Yoon Lee [email protected] 1
Department of BioNano Technology, Gachon University, 1342 Seongnam-daero, Sujeong-gu, Seongnam, Gyeonggi-do 13120, Korea
samples from the field set to the laboratory is a time-consuming process. During sample transfer, the condition should also be rigorously controlled to
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