Accurate Zygote‐Specific Discrimination of Single‐Nucleotide Polymorphisms Using Microfluidic Electrochemical DNA Melting Curves

Yang, Allen H. J.; Hsieh, Kuangwen; Patterson, Adriana S.; Ferguson, Blake; Eisenstein, Michael; Soh, H. Tom

doi:10.1002/ange.201310059

Cited by 13 publications

(10 citation statements)

References 39 publications

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“…A voltammetric sensor integrated within a microfluidic device was designed to perform electrochemical melting curve measurements and subsequently discriminate between homozygous and heterozygous SNPs in the apolipoprotein E (ApoE) gene. 414 Noteworthy is that specific alleles of ApoE strongly affect individual risk of late-onset Alzheimer's disease. The device employs methylene blue-labeled surface-bound DNA probes complementary to the SNP-containing target sequence.…”

Section: Electrochemical Detection Of Clinically Relevant Nucleic Acidsmentioning

confidence: 99%

Electrochemical Methods for the Analysis of Clinically Relevant Biomolecules

Sargent²,

2016

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Rapid progress in identifying biomarkers that are hallmarks of disease has increased demand for high-performance detection technologies. Implementation of electrochemical methods in clinical analysis may provide an effective answer to the growing need for rapid, specific, inexpensive, and fully automated means of biomarker analysis. This Review summarizes advances from the past 5 years in the development of electrochemical sensors for clinically relevant biomolecules, including small molecules, nucleic acids, and proteins. Various sensing strategies are assessed according to their potential for reaching relevant limits of sensitivity, specificity, and degrees of multiplexing. Furthermore, we address the remaining challenges and opportunities to integrate electrochemical sensing platforms into point-of-care solutions.

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Section: Electrochemical Detection Of Clinically Relevant Nucleic Acidsmentioning

confidence: 99%

Electrochemical Methods for the Analysis of Clinically Relevant Biomolecules

Sargent²,

2016

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“…30 Electrochemical sensing has been utilized for detection of single nucleic polymorphisms, 31 nucleic acids, 30d,32 and amplification products. 33 LAMP-based electrochemical sensing relies upon the oxidation–reduction of electroanalyte as monitored by linear sweep voltammetry (LSV), 34 square wave voltammetry (SWV), 33b,35 differential pulse voltammetry (DPV), 36 or conductivity.…”

Section: End-point Detectionmentioning

confidence: 99%

Emerging Loop-Mediated Isothermal Amplification-Based Microchip and Microdevice Technologies for Nucleic Acid Detection

Safavieh

Kanakasabapathy

Tarlan

et al. 2016

ACS Biomater. Sci. Eng.

155

104

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Rapid, sensitive, and selective pathogen detection is of paramount importance in infectious disease diagnosis and treatment monitoring. Currently available diagnostic assays based on polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA) are time-consuming, complex, and relatively expensive, thus limiting their utility in resource-limited settings. Loop-mediated isothermal amplification (LAMP) technique has been used extensively in the development of rapid and sensitive diagnostic assays for pathogen detection and nucleic acid analysis and hold great promise for revolutionizing point-of-care molecular diagnostics. Here, we review novel LAMP-based lab-on-a-chip (LOC) diagnostic assays developed for pathogen detection over the past several years. We review various LOC platforms based on their design strategies for pathogen detection and discuss LAMP-based platforms still in development and already in the commercial pipeline. This review is intended as a guide to the use of LAMP techniques in LOC platforms for molecular diagnostics and genomic amplifications.

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“…Performing multiple pathogen-specific reactions in parallel is one solution, but this approach can be inefficient and cost-prohibitive for large numbers of targets. Alternatively, one might employ universal primers to amplify sequences that can be subsequently differentiated via either sequence-specific probes (as demonstrated with our LAMP chip and elsewhere − ) or labeled with different redox reporters. , However, the extent to which this approach can be scaled up remains to be seen.…”

Section: Discussionmentioning

confidence: 99%

Integrated Electrochemical Microsystems for Genetic Detection of Pathogens at the Point of Care

et al. 2015

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The capacity to achieve rapid, sensitive, specific, quantitative, and multiplexed genetic detection of pathogens via a robust, portable, point-of-care platform could transform many diagnostic applications. And while contemporary technologies have yet to effectively achieve this goal, the advent of microfluidics provides a potentially viable approach to this end by enabling the integration of sophisticated multistep biochemical assays (e.g., sample preparation, genetic amplification, and quantitative detection) in a monolithic, portable device from relatively small biological samples. Integrated electrochemical sensors offer a particularly promising solution to genetic detection because they do not require optical instrumentation and are readily compatible with both integrated circuit and microfluidic technologies. Nevertheless, the development of generalizable microfluidic electrochemical platforms that integrate sample preparation and amplification as well as quantitative and multiplexed detection remains a challenging and unsolved technical problem. Recognizing this unmet need, we have developed a series of microfluidic electrochemical DNA sensors that have progressively evolved to encompass each of these critical functionalities. For DNA detection, our platforms employ label-free, single-step, and sequence-specific electrochemical DNA (E-DNA) sensors, in which an electrode-bound, redox-reporter-modified DNA "probe" generates a current change after undergoing a hybridization-induced conformational change. After successfully integrating E-DNA sensors into a microfluidic chip format, we subsequently incorporated on-chip genetic amplification techniques including polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP) to enable genetic detection at clinically relevant target concentrations. To maximize the potential point-of-care utility of our platforms, we have further integrated sample preparation via immunomagnetic separation, which allowed the detection of influenza virus directly from throat swabs and developed strategies for the multiplexed detection of related bacterial strains from the blood of septic mice. Finally, we developed an alternative electrochemical detection platform based on real-time LAMP, which not is only capable of detecting across a broad dynamic range of target concentrations, but also greatly simplifies quantitative measurement of nucleic acids. These efforts represent considerable progress toward the development of a true sample-in-answer-out platform for genetic detection of pathogens at the point of care. Given the many advantages of these systems, and the growing interest and innovative contributions from researchers in this field, we are optimistic that iterations of these systems will arrive in clinical settings in the foreseeable future.

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Accurate Zygote‐Specific Discrimination of Single‐Nucleotide Polymorphisms Using Microfluidic Electrochemical DNA Melting Curves

Cited by 13 publications

References 39 publications

Electrochemical Methods for the Analysis of Clinically Relevant Biomolecules

Electrochemical Methods for the Analysis of Clinically Relevant Biomolecules

Emerging Loop-Mediated Isothermal Amplification-Based Microchip and Microdevice Technologies for Nucleic Acid Detection

Integrated Electrochemical Microsystems for Genetic Detection of Pathogens at the Point of Care

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