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Separation and colorimetric detection of Escherichia coli by phage tail fiber protein combined with nano-magnetic beads

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Abstract

A colorimetric detection method for Escherichia coli (E. coli) in water was established based on a T7 phage tail fiber protein-magnetic separation. Firstly, the tail fiber protein (TFP) was expressed and purified to specifically recognize E. coli, which was verified by using fusion protein GFP-tagged TFP (GFP-TFP) and fluorescence microscopy. Then TFP conjugated with magnetic beads were applied to capture and separate E. coli. The TFP was covalently immobilized on the surface of magnetic beads and captured E. coli as verified by scanning electron microscopy (SEM). Finally, polymyxin B was used to lyse E. coli in solution and the released intracellular β-galactosidase (β-gal) could hydrolyze the colorimetric substrate chlorophenol red-β-D-galactopyranoside (CPRG), causing color change from yellow to purple. The high capture efficiencies of E. coli ranged from 88.70% to 95.65% and E. coli could be detected at a concentration of 102 CFU/mL by naked eyes. The specificity of the chromogenic substrate was evaluated using five different pathogen strains as competitors and tests with four kinds of real water samples showed recoveries of 86.00% to 92.25%. The colorimetric changes determined by visual inspection can be developed as an efficient platform for point-of-care detection of E. coli in resource-limited regions.

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Acknowledgements

This work was supported by Science and Technology Program of Guangzhou (202205110007).

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Authors and Affiliations

  1. School of Biology and Biological Engineering, South China University of Technology, Guangzhou, 510006, China

    Bin Hong, Yanmei Li, Wenhai Wang, Yi Ma & Jufang Wang

  2. Guangdong Provincial Key Laboratory of Fermentation and Enzyme Engineering, South China University of Technology, Guangzhou, China

    Yi Ma & Jufang Wang

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  1. Bin Hong
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  2. Yanmei Li
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  3. Wenhai Wang
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  4. Yi Ma
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Contributions

Bin Hong, Yi Ma and Jufang Wang, designed the experiments. Bin Hong, wrote the manuscript and performed the experiments. Yanmei Li, provided writing assistance for the manuscript. Wenhai Wang, analyzed data and drafted the manuscript. All authors have approved and read the final manuscript.

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Correspondence to Yi Ma or Jufang Wang.

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Highlights

1.A colorimetric detection method for E. coli in water was established based on a T7 phage tail fiber protein-magnetic separation.

2.Polymyxin B showed good lytic ability and was used to lyse E. coli to release β-gal.

3.There are high separation rate and low limit of detection for E. coli in our study.

4.The colorimetric change can be determined by naked eyes allowing for point-of-care testing of E. coli in resource-limited regions.

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Hong, B., Li, Y., Wang, W. et al. Separation and colorimetric detection of Escherichia coli by phage tail fiber protein combined with nano-magnetic beads. Microchim Acta 190, 202 (2023). https://doi.org/10.1007/s00604-023-05784-1

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