A label-free electrochemical biosensor for highly sensitive and selective detection of DNA via a dual-amplified strategy

Kong, Rongmei; Song, Zhiling; Meng, Hong-Min; Zhang, Xiaobing; Shen, Guo‐Li; Yu, Ru‐Qin

doi:10.1016/j.bios.2013.11.041

Cited by 63 publications

(13 citation statements)

References 39 publications

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“…Using the cascade signal amplification strategy of the HCR and the enzyme-induced metallization improved the detection sensitivity down to 10 pM, which is about 100-fold lower than that of the conventional unamplified homogeneous assays (Hu et al 2014). The detection sensitivity of this method could be compared to other amplification strategies (Gao and Li 2013; Kong et al 2014; Luo et al 2012; Zhang et al 2013), in which the detection limits were reported commonly in the range of fM to nM. For our method, when the concentration of the T DNA reached down to the 0.1–1 pM range, it was difficult to distinguish the colour change by the naked eyes.…”

Section: Resultsmentioning

confidence: 97%

“…Due to the low abundance of disease specific DNA and the complexity of the biological samples, highly sensitive and selective approaches for DNA detection are required in clinical applications. To meet this challenge, many methods have been well developed, for example, electrochemical sensing (Kong et al 2014; Liu et al 2013a), fluorescence (Hu et al 2013; Niu et al 2010) and chemiluminescence detection (Li and He 2009; Wang et al 2013c), and föster resonance energy resonance energy transfer (FRET) (Liu et al 2013b; Su et al 2014; Xing et al 2013). To further boost the performance, signal amplification strategies with various enzymes are usually adopted, such as exonuclease III-assisted amplification (Gao and Li 2013; Luo et al 2012), rolling circle amplification (Xu et al 2012), and strand displacement amplification (Wang et al 2011; Zhang et al 2013).…”

Section: Introductionmentioning

confidence: 99%

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Highly sensitive DNA detection using cascade amplification strategy based on hybridization chain reaction and enzyme-induced metallization

Zhang

Zheng

2015

Biosensors and Bioelectronics

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A novel highly sensitive colorimetric assay for DNA detection using cascade amplification strategy based on hybridization chain reaction and enzyme-induced metallization was established. The DNA modified superparamagnetic beads were demonstrated to capture and enrich the target DNA in the hybridization buffer or human plasma. The hybridization chain reaction and enzyme-induced silver metallization on the gold nanoparticles were used as cascade signal amplification for the detection of target DNA. The metalization of silver on the gold nanoparticles induced a significant colour change from red to yellow until black depending on the concentration of the target DNA, which could be recognized by naked eyes. This method showed a good specificity for the target DNA detection, with the capabilty to discriminate single-base-pair mismatched DNA mutation (single nucleotide polymorphism). Meanwhile, this approach exhibited an excellent anti-interference capability with the convenience of the magentic seperation and washing, which enabled its usage in complex biological systems such as human blood plasma. As an added benefit, the utilization of hybridization chain reaction and enzyme-induced metallization improved detection sensitivity down to 10 pM, which is about 100-fold lower than that of traditional unamplified homogeneous assays.

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Section: Resultsmentioning

confidence: 97%

Section: Introductionmentioning

confidence: 99%

Highly sensitive DNA detection using cascade amplification strategy based on hybridization chain reaction and enzyme-induced metallization

Zhang

Zheng

2015

Biosensors and Bioelectronics

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“…The regression equation was I ∆F = 105.1 logC + 1709 (R: 0.993), where ∆F = F–F0 (F: fluorescence signal of methylated DNA; F0: blank signal), I∆F is the relative fluorescence intensity and C is the methylated DNA concentration. Notably, LOD was 0.4 fM (based on 3σ/slope) [54]. As evidenced by both low LOD and wide linear range, this ultrasensitive fluorescent biosensor can be applied to detect low-level DNA methylation.…”

Section: Resultsmentioning

confidence: 99%

A novel fluorescent biosensor based on dendritic DNA nanostructure in combination with ligase reaction for ultrasensitive detection of DNA methylation

Zhang

Huang

et al. 2019

J Nanobiotechnol

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BackgroundDNA methylation detection is indispensable for the diagnosis and prognosis of various diseases including malignancies. Hence, it is crucial to develop a simple, sensitive, and specific detection strategy.MethodsA novel fluorescent biosensor was developed based on a simple dual signal amplification strategy using functional dendritic DNA nanostructure and signal-enriching polystyrene microbeads in combination with ligase detection reaction (LDR). Dendritic DNA self-assembled from Y-DNA and X-DNA through enzyme-free DNA catalysis of a hairpin structure, which was prevented from unwinding at high temperature by adding psoralen. Then dendritic DNA polymer labeled with fluorescent dye Cy5 was ligated with reporter probe into a conjugate. Avidin-labeled polystyrene microbeads were specifically bound to biotin-labeled capture probe, and hybridized with target sequence and dendritic DNA. LDR was triggered by adding Taq ligase. When methylated cytosine existed, the capture probe and reporter probe labeled with fluorescent dye perfectly matched the target sequence, forming a stable duplex to generate a fluorescence signal. However, after bisulfite treatment, unmethylated cytosine was converted into uracil, resulting in a single base mismatch. No fluorescence signal was detected due to the absence of duplex.ResultsThe obtained dendritic DNA polymer had a large volume. This method was time-saving and low-cost. Under the optimal experimental conditions using avidin-labeled polystyrene microbeads, the fluorescence signal was amplified more obviously, and DNA methylation was quantified ultrasensitively and selectively. The detection range of this sensor was 10−15 to 10−7 M, and the limit of detection reached as low as 0.4 fM. The constructed biosensor was also successfully used to analyze actual samples.ConclusionThis strategy has ultrasensitivity and high specificity for DNA methylation quantification, without requiring complex processes such as PCR and enzymatic digestion, which is thus of great value in tumor diagnosis and biomedical research.

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“…To date, various electrochemical DNA biosensors have been reported. To our knowledge, developing of label-free electrochemical DNA biosensors has been proved to be one successful method to directly detect target DNA [9][10][11]. As is well known, toluidine blue (TB) is a kind of phenothiazine dye and has good reversible electron transfer ability.…”

Section: Introductionmentioning

confidence: 99%

Label-free electrochemical DNA biosensor for rapid detection of mutidrug resistance gene based on Au nanoparticles/toluidine blue–graphene oxide nanocomposites

Peng

Liu

et al. 2015

Sensors and Actuators B: Chemical

146

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A label-free electrochemical biosensor for highly sensitive and selective detection of DNA via a dual-amplified strategy

Cited by 63 publications

References 39 publications

Highly sensitive DNA detection using cascade amplification strategy based on hybridization chain reaction and enzyme-induced metallization

Highly sensitive DNA detection using cascade amplification strategy based on hybridization chain reaction and enzyme-induced metallization

A novel fluorescent biosensor based on dendritic DNA nanostructure in combination with ligase reaction for ultrasensitive detection of DNA methylation

Label-free electrochemical DNA biosensor for rapid detection of mutidrug resistance gene based on Au nanoparticles/toluidine blue–graphene oxide nanocomposites

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