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Dual-mode aptasensor for simultaneous detection of multiple food-borne pathogenic bacteria based on colorimetry and microfluidic chip using stir bar sorptive extraction

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Abstract

A dual-mode aptasensor using colorimetry and microfluidic chip (MC) together with stir bar sorptive extraction (SBSE) has been developed for firstly qualifying samples contaminated with Vibrio parahaemolyticus (V.P) and Salmonella typhimurium (S.T), then precisely determine both of them in positive samples. For this purpose, the aptamer-streptavidin encoded probes (Apt-SAEs) corresponding to different bacteria were prepared in advance. Then, a stir bar modified with 4-mercaptophenylboronic acid (MPBA) was made to extract bacteria together with Apt-SAE probes. The binding event of aptamer and target triggered the formation of two sandwich structures containing Apt-SAE, V.P or S.T. The concentration of bacteria could be enriched by 1000 times within 15 min to avoid long-time enrichment process. Finally, the stir bar was immersed in the 3,3′,5,5′-Tetramethylbenzidine (TMB)-H2O2 solution for color development. The color could be observed by naked eyes to judge whether the analytes were present. The colorless samples were judged to be negative. For the positive samples, the adsorbed encoded probes corresponding to different bacteria would be eluted from the stir bar and rapidly analyzed by the MC. Under the optimized conditions, 100 CFU/mL of V.P or S.T or both of them could be observed by colorimetry and 35 CFU/mL of them could be detected (S/N = 3) by the MC. The assay has significant application value for on-site screening and multiple detection of food-borne pathogenic bacteria.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (21974074), Zhejiang Provincial Natural Science Foundation of China (LY19B050001, LY20B050004), Natural Science Foundation of Ningbo (2019A610184, 2019A610197), Zhejiang Province Public Welfare Technology Application Research Project (LGN18H300001, LGC20B050006, LGC19B070003), Zhejiang Provincial Top Discipline of Biological Engineering(Level A) (KF2019001), Basic Research Project of Key Laboratory of Guangzhou (202102100001) and K. C. Wong Magna Fund in Ningbo University.

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Authors and Affiliations

  1. Faculty of Material Science and Chemical Engineering, Ningbo University, Ningbo, 315211, China

    Ming Wang, Jin Zeng, Jiaqi Wang, Xiao Wang & Ning Gan

  2. Zhejiang Pharmaceutical College, Ningbo, 315211, China

    Ye Wang

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  1. Ming Wang
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  2. Jin Zeng
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Correspondence to Ye Wang or Ning Gan.

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Wang, M., Zeng, J., Wang, J. et al. Dual-mode aptasensor for simultaneous detection of multiple food-borne pathogenic bacteria based on colorimetry and microfluidic chip using stir bar sorptive extraction. Microchim Acta 188, 244 (2021). https://doi.org/10.1007/s00604-021-04902-1

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