Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Sensitive and Extraction-Free Detection of Methicillin-Resistant Staphylococcus aureus through Ag+ Aptamer-Based Color Reaction

  • Hongli Cao (Emergency Department, Beijing Rehabilitation Hospital, Capital Medical University) ;
  • Guosheng Zhang (Emergency Department, Beijing Rehabilitation Hospital, Capital Medical University) ;
  • Hui Ma (Emergency Department, Beijing Rehabilitation Hospital, Capital Medical University) ;
  • Zhongwen Xue (Emergency Department, Beijing Rehabilitation Hospital, Capital Medical University) ;
  • Ran Huo (Emergency Department, Beijing Rehabilitation Hospital, Capital Medical University) ;
  • Kun Wang (Emergency Department, Beijing Rehabilitation Hospital, Capital Medical University) ;
  • Zijin Liu (Orthopedic Rehabilitation Department, Beijing Rehabilitation Hospital, Capital Medical University)
  • Received : 2023.08.24
  • Accepted : 2023.09.14
  • Published : 2024.01.28
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Abstract

Refractory infections, such as hospital-acquired pneumonia, can be better diagnosed with the assistance of precise methicillin-resistant Staphylococcus aureus (MRSA) testing. However, traditional methods necessitate high-tech tools, rigorous temperature cycling, and the extraction of genetic material from MRSA cells. Herein, we propose a sensitive, specific, and extraction-free strategy for MRSA detection by integrating allosteric probe-based target recognition and exonuclease-III (Exo-III)-enhanced color reaction. The penicillin-binding protein 2a (PBP2a) aptamer in the allosteric probe binds with MRSA to convert protein signals to nucleic acid signals. This is followed by the DNA polymerase-assisted target recycle and the production of numerous single-strand DNA (ssDNA) chains which bind with silver ion (Ag+) aptamer to form a blunt terminus that can be identified by Exo-III. As a result, the Ag+ aptamer pre-coupled to magnetic nanoparticles is digested. After magnetic separation, the Ag+ in liquid supernatant catalyzes 3,3',5,5'-tetramethylbenzidine (TMB) for a color reaction. In addition, a concentration of 54 cfu/mL is predicted to be the lowest detectable value. Based on this, our assay has a wide linear detection range, covering 5 orders of magnitude and demonstrating a high specificity, which allows it to accurately distinguish the target MRSA from other microorganisms.

Keywords

Acknowledgement

This work was supported by the First Affiliated Hospital of Capital Medical University.

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