A versatile label-free electrochemical biosensor for circulating tumor DNA based on dual enzyme assisted multiple amplification strategy

Wang, Huafeng; Ma, Rong-Na; Sun, Fei; Jia, Liping; Zhang, Wei; Shang, Lei; Xue, Qingwang; Jia, Wen-Li

doi:10.1016/j.bios.2018.09.028

Cited by 75 publications

(33 citation statements)

References 37 publications

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“…In addition to the evaluation of potential measurement and signal amplification schemes, the importance of the underlying electrode has also been investigated through the use of e.g., nanostructured gold electrodes [18]. What these advances have so far reported are complex assays involving the use of modified oligonucleotides [19], formation of branched structures upon DNA recognition [20], molecular switches [21], nanostructured electrode modifications [18], etc., therefore ultimately having a limited chance of manufacture and clinical uptake. The current state-of-the-art in terms of general nucleic acid liquid biopsies [22] and electrochemical detection of ctDNA biomarkers [23] have been well-reviewed recently.…”

Section: Introductionmentioning

confidence: 99%

Developing a Low-Cost, Simple-to-Use Electrochemical Sensor for the Detection of Circulating Tumour DNA in Human Fluids

et al. 2020

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It is well-known that two major issues, preventing improved outcomes from cancer are late diagnosis and the evolution of drug resistance during chemotherapy, therefore technologies that address these issues can have a transformative effect on healthcare workflows. In this work we present a simple, low-cost DNA biosensor that was developed specifically to detect mutations in a key oncogene (KRAS). The sensor employed was a screen-printed array of carbon electrodes, used to perform parallel measurements of DNA hybridisation. A DNA amplification reaction was developed with primers for mutant and wild type KRAS sequences which amplified target sequences from representative clinical samples to detectable levels in as few as twenty cycles. High levels of sensitivity were demonstrated alongside a clear exemplar of assay specificity by showing the mutant KRAS sequence was detectable against a significant background of wild type DNA following amplification and hybridisation on the sensor surface. The time to result was found to be 3.5 h with considerable potential for optimisation through assay integration. This quick and versatile biosensor has the potential to be deployed in a low-cost, point-of-care test where patients can be screened either for early diagnosis purposes or monitoring of response to therapy.

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Section: Introductionmentioning

confidence: 99%

Developing a Low-Cost, Simple-to-Use Electrochemical Sensor for the Detection of Circulating Tumour DNA in Human Fluids

et al. 2020

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“…[30] In one example,atriplehelix molecular switch on the sensor is opened by target ctDNA. [30] In one example,atriplehelix molecular switch on the sensor is opened by target ctDNA.…”

Section: Methodsmentioning

confidence: 99%

High‐Performance Nucleic Acid Sensors for Liquid Biopsy Applications

Das,

Kelley

2019

Angew Chem Int Ed

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Circulating tumour nucleic acids (ctNAs) are released from tumours cells and can be detected in blood samples, providing a way to track tumors without requiring a tissue sample. This “liquid biopsy” approach has the potential to replace invasive, painful, and costly tissue biopsies in cancer diagnosis and management. However, a very sensitive and specific approach is required to detect relatively low amounts of mutant sequences linked to cancer because they are masked by the high levels of wild‐type sequences. This review discusses high‐performance nucleic acid biosensors for ctNA analysis in patient samples. We compare sequencing‐ and amplification‐based methods to next‐generation sensors for ctDNA and ctRNA (including microRNA) profiling, such as electrochemical methods, surface plasmon resonance, Raman spectroscopy, and microfluidics and dielectrophoresis‐based assays. We present an overview of the analytical sensitivity and accuracy of these methods as well as the biological and technical challenges they present.

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“…Testing this mutation, among others, codon 12 and 13 in the KRAS gene, is now put in practice [39,40]. Hua-Feng Wang et al [106] developed an ultrasensitive label-free biosensor based in a dual enzyme-assisted multiple amplification (RNase HII: Ribonuclease HII, and TdT: terminal deoxynucleotidyl transferase). The principle of the biosensor is schematically illustrated in Figure 2 and consists of three key steps.…”

Section: Electrochemical Biosensing Of Nucleic Acid Biomarkers Of Crcmentioning

confidence: 99%

mentioning

confidence: 99%

Electrochemical Biosensors for Determination of Colorectal Tumor Biomarkers

et al. 2020

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The accurate determination of specific tumor markers associated with cancer with non-invasive or minimally invasive procedures is the most promising approach to improve the long-term survival of cancer patients and fight against the high incidence and mortality of this disease. Quantification of biomarkers at different stages of the disease can lead to an appropriate and instantaneous therapeutic action. In this context, the determination of biomarkers by electrochemical biosensors is at the forefront of cancer diagnosis research because of their unique features such as their versatility, fast response, accurate quantification, and amenability for multiplexing and miniaturization. In this review, after briefly discussing the relevant aspects and current challenges in the determination of colorectal tumor markers, it will critically summarize the development of electrochemical biosensors to date to this aim, highlighting the enormous potential of these devices to be incorporated into the clinical practice. Finally, it will focus on the remaining challenges and opportunities to bring electrochemical biosensors to the point-of-care testing.

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A versatile label-free electrochemical biosensor for circulating tumor DNA based on dual enzyme assisted multiple amplification strategy

Cited by 75 publications

References 37 publications

Developing a Low-Cost, Simple-to-Use Electrochemical Sensor for the Detection of Circulating Tumour DNA in Human Fluids

Developing a Low-Cost, Simple-to-Use Electrochemical Sensor for the Detection of Circulating Tumour DNA in Human Fluids

High‐Performance Nucleic Acid Sensors for Liquid Biopsy Applications

Electrochemical Biosensors for Determination of Colorectal Tumor Biomarkers

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