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Application of a cation-exchange reaction of CuS nanoparticles and fluorescent copper nanoparticles in a DNA biosensor.

ABSTRACT: A novel detection method based on the cation-exchange reaction of CuS nanoparticles (CuS NPs) combined with poly T-templated fluorescent Cu nanoparticles (Cu NPs) was developed. First, CuS NPs-magnetic bead conjugates were prepared through the hybridization of DNA. Competition with target DNA resulted in the release of CuS NPs, and exonuclease III catalysis could lead to recycling of the target DNA. Then, the CuS NPs released into the supernatant were subjected to a cation-exchange reaction after the addition of AgNO₃. The obtained Cu²⁺ could form fluorescent Cu NPs using poly T DNA as a template. The fluorescence intensity of the Cu NPs could be used to determine the concentration of the target DNA. To further increase the detection sensitivity, two types of DNA decorated magnetic beads were used. After Exo III digestion for two cycle processes, more CuS NPs entered the supernatant. Hence, a stronger fluorescence intensity was found after the cation-exchange reaction and the formation of fluorescent Cu NPs. The developed method is convenient and low cost with good sensitivity and selectivity.

SUBMITTER: Song W

PROVIDER: S-EPMC9080010 | biostudies-literature | 2018 Apr

REPOSITORIES: biostudies-literature

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Application of a cation-exchange reaction of CuS nanoparticles and fluorescent copper nanoparticles in a DNA biosensor.

Song Weiling W Zhang Nan N Luan Zhenzhu Z Zhang Xiaoru X He Peng P

RSC advances 20180423 27

A novel detection method based on the cation-exchange reaction of CuS nanoparticles (CuS NPs) combined with poly T-templated fluorescent Cu nanoparticles (Cu NPs) was developed. First, CuS NPs-magnetic bead conjugates were prepared through the hybridization of DNA. Competition with target DNA resulted in the release of CuS NPs, and exonuclease III catalysis could lead to recycling of the target DNA. Then, the CuS NPs released into the supernatant were subjected to a cation-exchange reaction afte ...[more]

PMID: 35541366

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Project description:BACKGROUND:DNA 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. METHODS:A 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. RESULTS:The 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. CONCLUSION:This 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.

Dataset Information

Application of a cation-exchange reaction of CuS nanoparticles and fluorescent copper nanoparticles in a DNA biosensor.

Publications

Application of a cation-exchange reaction of CuS nanoparticles and fluorescent copper nanoparticles in a DNA biosensor.

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