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Sensitive and Extraction-Free Detection of Methicillin-Resistant Staphylococcus aureus through Ag+ Aptamer-Based Color Reaction.


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.

SUBMITTER: Cao H 

PROVIDER: S-EPMC10840478 | biostudies-literature | 2024 Jan

REPOSITORIES: biostudies-literature

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Sensitive and Extraction-Free Detection of Methicillin-Resistant <i>Staphylococcus aureus</i> through Ag<sup>+</sup> Aptamer-Based Color Reaction.

Cao Hongli H   Zhang Guosheng G   Ma Hui H   Xue Zhongwen Z   Huo Ran R   Wang Kun K   Liu Zijin Z  

Journal of microbiology and biotechnology 20230928 1


Refractory infections, such as hospital-acquired pneumonia, can be better diagnosed with the assistance of precise methicillin-resistant <i>Staphylococcus aureus</i> (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-II  ...[more]

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