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Ultrasensitive colorimetric detection of Staphylococcus aureus using wheat germ agglutinin and IgY as a dual-recognition strategy

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Abstract

A novel colorimetric platform was designed for the determination of S. aureus by utilizing a dual-recognition strategy, where wheat germ agglutinin (WGA)-functionalized magnetic beads were served as separation elements to capture and enrich S. aureus efficiently from the matrix. Horseradish peroxidase (HRP) labeled chicken anti-protein A IgY (HRP-IgY) was used to label the captured S. aureus. A chicken IgY was introduced as a signal tracer to bind with staphylococcal protein A (SPA) on the surface of S. aureus, which can circumvent the interference from protein G-producing Streptococcus. Subsequently, the colorimetric signal was achieved by an HRP-catalyzed reaction, which was amplified by HRP-IgY bound by approximately 80,000 SPA molecules on one S. aureus. The entire detection process could be accomplished within 90 min. Under optimal conditions, the linear response of different S. aureus concentrations ranged from 7.8 × 102 to 2.0 × 105 CFU/mL and the limit of detection reached down to 3.9 × 102 CFU/mL. Some common non-target bacteria yielded negative results, indicating the excellent specificity of the method. The developed strategy was successfully applied to the determination of S. aureus in various types of samples with satisfactory recoveries. Therefore, the novel dual-recognition strategy possessed the advantages of high sensitivity, specificity, and low cost and exhibited considerable potential as a promising tool to defend public health.

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Funding

This work was funded by the National Natural Science Foundation of China (Grant No. 81702099), Key Scientific Research Projects of Key Scientific and Technological Projects of Henan Province (Nos. 222102310476 and 222102310310), and Key Scientific Research Projects of Higher Education Institutions in Henan Province (Grant No. 23A330004).

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Authors and Affiliations

  1. Laboratory of Infection and Immunology, School of Medical Technology, Xinxiang Medical University, Xinxiang, 453003, People’s Republic of China

    Yun Zhang, Ge Tian, Xueni Sun, Xiaoli Yang, Yi Zhang, Wenqing Tan, Liangwei Duan & Junping Yu

  2. CAS Key Laboratory of Special Pathogens and Biosafety, Center for Emerging Infectious Diseases, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, 430071, People’s Republic of China

    Junping Yu

  3. Department of Ophthalmology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200080, People’s Republic of China

    Shunxiang Gao

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  1. Yun Zhang
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  2. Ge Tian
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  3. Xueni Sun
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  4. Xiaoli Yang
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  5. Yi Zhang
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  7. Liangwei Duan
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  9. Junping Yu
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Contributions

YZ: conceptualization, methodology, investigation, and writing—original draft. GT: investigation and methodology. XS: investigation and formal analysis. XY: investigation and methodology. YZ: validation, software, and data curation. WT: methodology. LD: formal analysis. SG: conceptualization and validation. JY: resources, writing—review and editing, supervision, and project administration.

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Correspondence to Liangwei Duan, Shunxiang Gao or Junping Yu.

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Zhang, Y., Tian, G., Sun, X. et al. Ultrasensitive colorimetric detection of Staphylococcus aureus using wheat germ agglutinin and IgY as a dual-recognition strategy. Microchim Acta 191, 209 (2024). https://doi.org/10.1007/s00604-024-06288-2

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