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Exo-III Enzyme–Assisted Triple Cycle Signal Amplifications for Sensitive and Accurate Identification of Pathogenic Bacteria

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Abstract

Early determination of infectious pathogens is vitally important to select appropriate antibiotics, and to manage nosocomial infection. Herein, we propose a target recognition triggered triple signal amplification–based approach for sensitive pathogenic bacteria detection. In the proposed approach, a double-strand DNA probe (capture probe) that is composed of an aptamer sequence and a primer sequence is designed for specific identification of target bacteria and initiation of following triple signal amplification. After recognition of target bacteria, primer sequence is released from capture probe to bind with the designed H1 probe, forming a blunt terminal in the H1 probe. Exonuclease-III (Exo-III enzyme) specifically recognizes the blunt terminal in H1 probe and degrades the sequence from 3′ terminal, resulting a single-strand DNA to induce the following signal amplification. Eventually, the approach exhibits a low detection limit of 36 cfu/mL with a broad dynamic range. The high selectivity endows the method a promising prospective for clinical sample analysis.

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Authors and Affiliations

  1. Department of Endoscopy Center, Huangshi Central Hospital, Affiliated Hospital of Hubei Polytechnic University, Hubei Province, Huangshi, People’s Republic of China

    Jie Guo & Miao Tian

  2. Health Commission of Huangshi, Hubei Province, Huangshi, People’s Republic of China

    Qun Liang

  3. Department of Gastroenterology, Huangshi Central Hospital, Affiliated Hospital of Hubei Polytechnic University, Hubei Province, Huangshi, People’s Republic of China

    Huifang Zhang

  4. Department of Operation Management, Huangshi Central Hospital, Affiliated Hospital of Hubei Polytechnic University, Hubei Province, Huangshi, People’s Republic of China

    Huajun Zhang & Wantao Zhang

  5. Department of Pediatric, Huangshi Central Hospital, Affiliated Hospital of Hubei Polytechnic University, Huangshi, Hubei, People’s Republic of China

    Guo Wei

Authors
  1. Jie Guo
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  2. Qun Liang
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  3. Huifang Zhang
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  4. Miao Tian
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  5. Huajun Zhang
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  6. Guo Wei
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  7. Wantao Zhang
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Contributions

All authors contributed to the study conception and design. Jie Guo and Qun Liang coordinated all the experiments and wrote the manuscript draft. Huajun Zhang, Guo Wei, and Wantao Zhang designed the approach and revised the manuscript. Huifang Zhang and Miao Tian analyzed the research data and reviewed the manuscript. All authors have read and approved the final manuscript. Jie Guo, Qun Liang, and Huifang Zhang contributed equally to this work

Corresponding authors

Correspondence to Huajun Zhang, Guo Wei or Wantao Zhang.

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Guo, J., Liang, Q., Zhang, H. et al. Exo-III Enzyme–Assisted Triple Cycle Signal Amplifications for Sensitive and Accurate Identification of Pathogenic Bacteria. Appl Biochem Biotechnol 195, 6203–6211 (2023). https://doi.org/10.1007/s12010-023-04391-3

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  • DOI: https://doi.org/10.1007/s12010-023-04391-3

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