Helicase dependent OnChip-amplification and its use in multiplex pathogen detection

Andresen, Dennie; Nickisch‐Rosenegk, Markus von; Bier, Frank F.

doi:10.1016/j.cca.2009.03.021

Cited by 60 publications

(44 citation statements)

References 16 publications

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“…One primer was immobilized to the surface of an epoxysilanized glass slide. The fluorescently-labelled second primer and the sample containing HDA enzymes were incubated for two hours to allow for bacterial pathogen monitoring [114]. The detection of Staphylococcus aureus from nasal swabs was presented in a study from Frech et al, were an asymmetric HDA was used to amplify specific sequences, which hybridized to capture probes on a DNA array.…”

Section: E Colimentioning

confidence: 99%

Isothermal Amplification and Quantification of Nucleic Acids and its Use in Microsystems

Tröger¹,

Niemann²,

Gärtig³

et al. 2015

J Nanomed Nanotechnol

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Nucleic acid amplification technologies (NAATs) offer the most sensitive tests in the clinical laboratory. These techniques are used as a powerful tool for screening and diagnosis of infectious diseases. Isothermal methods, as an alternative to polymerase chain reaction (PCR), require no thermocycling machine and can mostly be performed with reduced time, high throughput, and accurate and reliable results.However, current molecular diagnostic approaches generally need manual analysis by qualified and experienced personal which is a highly complex, time-consuming and labor-intensive task. Thus, the demand for simpler, miniaturized systems and assays for pathogen detection is steadily increasing. Microfluidic platforms and lab-on-a-chip devices have many advantages such as small sample volume, portability and rapid detection time and enable point-of-care diagnosis.In this article, we review several isothermal amplification methods and their implementation in microsystems in relation to quantification of nucleic acids. miniaturized systems can be diverse. The majority of publications are focussed on clinical diagnostics including foodborne organisms for environmental monitoring [10][11][12]. In case of an infectious disease, cultivation and phenotypic characterization are still the standard methods of microbiologists which need days up to weeks to obtain a diagnostic result. Hence, scientists started to make use of nucleic acid amplification methods to accelerate the analysis. The most widespread and well known technique for this purpose is the polymerase chain reaction (PCR) [10,13], which exploits the activity of the DNA polymerase. The method is based on a thermal cycling process consisting of repeated cycles of heating and cooling steps allowing for DNA melting, sequence specific primer binding and enzymatic amplification of the target nucleic acid. The quantitative assessment of target copies is possible by using a real-time PCR approach, where a concentration dependent signal (e.g. fluorescence) increases over time [14]. The latter enables the user to easily assess the pathogen load of patient samples [15][16][17][18]. Since the first example of a PCR on a miniaturized system in 1994 [19,20], numerous devices were presented, which integrate not just the amplification, but also sample preparation and detection steps [9,[21][22][23][24][25][26][27][28]. Just a few of those microsystems have reached the market so far, but some have shown a fascinating potential to replace the classical laboratory-oriented state-of-the art [29][30][31][32][33]. setups has been shown to obtain a similar result [34]. A smaller heat capacity allows for rapid changes in temperature beneficial for the PCR time as well as a higher parallelism of multiple genetic samples [34].However, a major drawback of the integration of PCR to microsystems is the necessity of sophisticated instrumentation. The reaction requires a thermal cycling instrumentation, space and considerable expertise [35]. Therefore, isothermal reactions for the ta...

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Section: E Colimentioning

confidence: 99%

Isothermal Amplification and Quantification of Nucleic Acids and its Use in Microsystems

Tröger¹,

Niemann²,

Gärtig³

et al. 2015

J Nanomed Nanotechnol

View full text Add to dashboard Cite

show abstract

“…When combined with microarray technology isothermal amplification can potentially open up new horizons as a low-cost, highly sensitive diagnostic tool for home use and/or personalized medicine. In particular, rolling circle amplification (RCA) [54], loop mediated isothermal amplification (LAMP) [55], helicase dependent amplification (HDA) [56] and strand displacement amplification [57][58][59]) are the most widely used techniques for isothermal DNA/RNA amplification. RCA is one of the simplest isothermal amplification processes in which one single-strand primer is extended by DNA polymerase with strand displacement activity using a circular DNA as a template.…”

Section: Isothermal Amplification On Microarraysmentioning

confidence: 99%

“…By utilizing primers immobilized on microbeads attomole detection of target circular DNA has been reported [62,63]. The use of LAMP [55] and HDA [56] methods on solid supports has also been successfully shown.…”

Section: Isothermal Amplification On Microarraysmentioning

confidence: 99%

“…Isothermal detection of methicillin resistant S. aureus (MRSA) and N. gonorrhoeae dsDNA has been carried out using HDA [56]. HDA is very similar to PCR amplification with the exception that HDA is performed at a constant temperature.…”

Section: Isothermal Amplification On Microarraysmentioning

confidence: 99%

“…A year later the same authors [56] reported on a further improvement of the approach and development of the ready-to-use cassette containing short glass capillaries filled with desiccated (for shelf storage) acrylamide gels with all the reagents required for the PCR reaction except the DNA template. In order to perform the nucleic acids testing reaction the cassette has to be filled with raw sample containing the target templates and thermo-cycled.…”

Section: Microfluidic Arraysmentioning

confidence: 99%

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Recent developments in nucleic acid identification using solid-phase enzymatic assays

Khodakov

Ellis

2014

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DNA‐Based Chemical Reaction Networks for Biosensing Applications

Jiang

Yuan

et al. 2020

Advanced Intelligent Systems

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The nature of biosensing is a biochemical reaction. DNA‐based chemical reaction networks (DNA‐CRNs) as a powerful programming language for describing behaviors of chemical reactions have shown great potential in designs and applications of biosensors. Due to their programmability, modularity, and versatility of the DNA strand, the performance of different detection strategies can be improved mainly by the rational design of DNA‐CRNs. Herein, an overview of the fundamental theory and biosensing processes of DNA‐CRNs is provided. Various detection strategies of DNA‐CRNs are introduced, either in a simple low‐order reaction model or in a complicated high‐order reaction type, in combination with some typical cases for the different detection purposes. In addition, an overview of the recent development of DNA‐CRNs for monitoring the cell microenvironment is presented, which is of significance to uncover some specific cell behaviors and functions. Finally, the roles of DNA‐CRNs in the rational design of high‐performance biosensors are summarized by pointing out the remaining challenges that impede the precise biosensing using DNA‐CRNs in complicated biological environments.

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Helicase dependent OnChip-amplification and its use in multiplex pathogen detection

Cited by 60 publications

References 16 publications

Isothermal Amplification and Quantification of Nucleic Acids and its Use in Microsystems

Isothermal Amplification and Quantification of Nucleic Acids and its Use in Microsystems

Recent developments in nucleic acid identification using solid-phase enzymatic assays

DNA‐Based Chemical Reaction Networks for Biosensing Applications

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