An efficient DNAzyme-based DNA scaffold for label-free and sensitive bacterial pathogen detection

Zhang, Xue; Zhang, Jiali; Wen, Yongbin

doi:10.1016/j.ab.2023.115076

Cited by 2 publications

(1 citation statement)

References 23 publications

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“…Therefore, one popular strategy with regard to electrochemical biosensors has been to modify signaling molecules, for example by conjugating them to MB or Fc, to achieve oligonucleotide-assisted ratiometric sensing. [24][25][26] The G-quadruplex is a special secondary structure of nucleic acid whose unique structure can combine a variety of signaling molecules, including electrochemical signal indicators such as MB, fluorophore thioflavin T (ThT), and others, [27,28] to achieve more ideal signal amplification effect and more sensitive detection. To obtain a large number of MB carriers in a Gquadruplex, rolling circle amplification (RCA), a common exponential amplification method, [29][30][31][32] can be supplemented to generate the majority of G-rich single-stranded oligonucleotide sequences for bonding to MB molecules.…”

Section: Introductionmentioning

confidence: 99%

Rolling Circle Amplification/G‐Quadruplex‐Based Dual‐Signal Ratiometric Electrochemical Aptasensor for Ultrasensitive Detection of Pathogenic Bacteria

et al. 2023

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Accurate and efficient detection of pathogenic bacteria plays a crucial role in the diagnosis of infectious diseases. However, rapid and highly sensitive detection of specific bacteria in clinical samples remains challenging. In this study, we developed a ratiometric dual‐signal electrochemical biosensor for ultrasensitive detection of pathogenic bacteria, based on an aptamer recognition‐induced rolling circle amplification (RCA)/G‐quadruplex strategy. On the surface of a gold electrode chip, we used an aptamer sequence (P1) conjugated to ferrocene (Fc) to capture the target bacterium. This capture released a different, previously bound sequence (P2) sequence that initiated the RCA/G‐quadruplex cascade, which, in the presence of potassium ions, was able to bind the electrochemical indicator methylene blue (MB). We integrated the signals from the loss of Fc and the appearance of MB (IMB/IFC ratio) to quantify the target bacteria concentration. This biosensor showed excellent detection performance and specificity, with a detection limit of 10 CFU/mL. Notably, it showed great diagnostic potential for clinical infectious diseases caused by pathogenic bacteria in precise/personalized medicine applications.

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Section: Introductionmentioning

confidence: 99%

Rolling Circle Amplification/G‐Quadruplex‐Based Dual‐Signal Ratiometric Electrochemical Aptasensor for Ultrasensitive Detection of Pathogenic Bacteria

et al. 2023

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Enhanced Detection of Vibrio harveyi Using a Dual-Composite DNAzyme-Based Biosensor

Li,

Zhang,

Jiang

et al. 2024

Biosensors

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Vibrio harveyi is a serious bacterial pathogen which can infect a wide range of marine organisms, such as marine fish, invertebrates, and shrimp, in aquaculture, causing severe losses. In addition, V. harveyi can be transmitted through food and water, infecting humans and posing a serious threat to public safety. Therefore, rapid and accurate detection of this pathogen is key for the prevention and control of related diseases. In this study, nine rounds of in vitro screening were conducted with Systematic Evolution of Ligands by Exponential Enrichment (SELEX) technology using unmodified DNA libraries, targeting the crude extracellular matrix (CEM) of V. harveyi. Two DNAzymes, named DVh1 and DVh3, with high activity and specificity were obtained. Furthermore, a fluorescent biosensor with dual DNAzymes was constructed which exhibited improved detection efficiency. The sensor showed a good fluorescence response to multiple aquatic products (i.e., fish, shrimp, and shellfish) infected with V. harveyi, with a detection limit below 11 CFU/mL. The fluorescence signal was observed within 30 min of reaction after target addition. This simple, inexpensive, highly effective, and easy to operate DNAzymes biosensor can be used for field detection of V. harveyi.

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An efficient DNAzyme-based DNA scaffold for label-free and sensitive bacterial pathogen detection

Cited by 2 publications

References 23 publications

Rolling Circle Amplification/G‐Quadruplex‐Based Dual‐Signal Ratiometric Electrochemical Aptasensor for Ultrasensitive Detection of Pathogenic Bacteria

Rolling Circle Amplification/G‐Quadruplex‐Based Dual‐Signal Ratiometric Electrochemical Aptasensor for Ultrasensitive Detection of Pathogenic Bacteria

Enhanced Detection of Vibrio harveyi Using a Dual-Composite DNAzyme-Based Biosensor

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