Sensitive and direct electrochemical detection of double-stranded DNA utilizing alkaline phosphatase-labelled zinc finger proteins

Noh, Soodong; Ha, Dat Thinh; Yang, Haesik; Kim, Moon‐Soo

doi:10.1039/c5an00623f

Cited by 22 publications

(26 citation statements)

References 34 publications

(68 reference statements)

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“…As demonstrated previously 9–11 as well as in this study, the use of ZFPs may lead to significant contribution to application of DNA-binding domains for developing new technology for pathogen detection. ZFP construction by modular assembly described in this study is relatively easy, thus the use of ZFPs for a real-world diagnostic can be cost-effective.…”

Section: Resultssupporting

confidence: 72%

“…11 Zn 2+ -containing HEPES buffer was used to maintain the ZFP structure during chemical conjugation. 11 500 µL of HEPES buffer containing 200 µg mL −1 ZFP was mixed with 10 µL of 4 mM EZ-link sulfo-NHS-LC-LC-biotin and incubated for 2 h at 4 °C, and the mixture was then filtered by centrifugation for 20 min. Afterward, the filtrate was dissolved in 1 mL of TZ buffer.…”

Section: Methodsmentioning

confidence: 99%

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Multiplexed detection of pathogen-specific DNA using engineered zinc finger proteins without target amplification

Kim

2016

Anal. Methods

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Multiplexed detection of pathogen-specific DNA sequences in a simple and reliable way is in great demand for clinical and biomedical applications. However, there is still a lack of available DNA detection methods that are simple and pathogen-selective for point-of-care (POC) testing. Here, we report a novel zinc finger protein (ZFP)-based chemiluminescent method for direct detection of pathogenic double-stranded DNA (dsDNA) in a multiplexed platform. ZFPs are custom-designed to identify unique pathogenic DNA sequences. ZFP-based chemiluminescent detection of dsDNA provides sufficient sensitivity (≤50 fmol) and high specificity without target DNA amplification. Our study addresses the potential of developing a simple and selective pathogen detection method in a multiplexed fashion needed for POC application.

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Section: Resultssupporting

confidence: 72%

Section: Methodsmentioning

confidence: 99%

Multiplexed detection of pathogen-specific DNA using engineered zinc finger proteins without target amplification

Kim

2016

Anal. Methods

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“…Many hybridization-based DNA detection methods use ssDNA as target DNA. However, the cfDNA present in the blood is dsDNA, and the DNA amplified by PCR is also dsDNA [188,189]. Therefore, a method for detecting dsDNA should be considered.…”

Section: Perspective On Nucleic Acid Detection In a Sample-to-answer Systemmentioning

confidence: 99%

Total microfluidic platform strategy for liquid biopsy

Lee

Shin

2021

JCB

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A liquid biopsy is a simple and non-invasive biopsy that examines a range of information about a tumor through a simple blood sample. Due to its non-invasive nature, liquid biopsy has many outstanding clinical benefits, including repetitive sampling and examination, representation of whole mutations, observation of minimal residual disease etc. However, liquid biopsy requires various processes such as sample preparation, amplification, and target detection. These processes can be integrated onto microfluidic platforms, which may provide a sample-to-answer system. The present review provides a brief overview of liquid biopsies, a detailed review of the technologies in each process, and prospective concluding remarks. Through this review, one can have a basic but cross-disciplinary understanding of liquid biopsy, as well as knowledge of new starting points for future research in each related area.

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“…In fact, DNA is rarely present in single-stranded form, either naturally or after PCR amplification. The improvements in the detection of dsDNA or natural DNA leads to more robust and flexible DNA diagnostics [10].…”

Section: Introductionmentioning

confidence: 99%

Ultrasensitive direct detection of dsDNA using a glassy carbon electrode modified with thionin-functionalized multiple graphene aerogel and gold nanostars

et al. 2016

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The paper reports on the synthesis of a composite consisting of thionin-functionalized multiple graphene aerogel and gold nanostars (termed TF-MGA/GNSs). The composite displays an enhanced electrochemical performance compared to the classical graphene aerogel. The performance can be further improved by increasing number of the graphene oxide gelation cycles. The composite was deposited on a glassy carbon electrode to obtain a biosensor for the direct detection of dsDNA. The interaction of thionin with dsDNA causes a specific electrochemical response, best at a working voltage of −228 mV (vs. Ag/AgCl). The enhanced electrocatalytic activity of the composite strongly improves sensitivity. The differential pulse voltammetric signal linearly decreases with increasing dsDNA concentration in the range from 100 fg · mL −1 to 10 ng · mL −1 , and the detection limit is 39 fg · mL −1 (at an S/N ratio of 3). The method is sensitive, selective and rapid. It was successfully applied to the detection of circulating free DNA in human serum. The study also provides an attractive approach towards graphene aerogel-based materials to meet the needs of nanoelectronics for molecule diagnosis, bioanalysis and catalysis.

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Sensitive and direct electrochemical detection of double-stranded DNA utilizing alkaline phosphatase-labelled zinc finger proteins

Cited by 22 publications

References 34 publications

Multiplexed detection of pathogen-specific DNA using engineered zinc finger proteins without target amplification

Multiplexed detection of pathogen-specific DNA using engineered zinc finger proteins without target amplification

Total microfluidic platform strategy for liquid biopsy

Ultrasensitive direct detection of dsDNA using a glassy carbon electrode modified with thionin-functionalized multiple graphene aerogel and gold nanostars

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