PMMA biosensor for nucleic acids with integrated mixer and electrochemical detection

Nugen, Sam R.; Asiello, Peter J.; Connelly, John T.; Baeumner, Antje J.

doi:10.1016/j.bios.2008.12.025

Cited by 86 publications

(68 citation statements)

References 39 publications

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“…102 In 2012, Agrawal and coworkers showed simultaneous detection of E. coli and S. tyhimurium on a circular microfluidics design manufactured in PDMS using magnetic nanoparticles to control and concentrate samples. 61 With regard to protozoa, two publications that relate to molecular sensing of Cryptosporidium in miniaturized format describe the performance of NASBA off-chip 103 ; only the detection of the mRNAs amplicons was performed on-chip. Esch et al have developed a fluorescence-based detection assay chip, relying on a sandwich hybridization of the NASBA product between capture probes and reporter probes.…”

Section: Waterborne Pathogensmentioning

confidence: 99%

Miniaturized Detection Systems

Bridle¹

2014

Waterborne Pathogens

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Section: Waterborne Pathogensmentioning

confidence: 99%

Miniaturized Detection Systems

Bridle¹

2014

Waterborne Pathogens

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“…The isothermal amplification enabled them to detect a single oocyst in a reaction volume of 4 µL [88].…”

Section: Nasba: Nucleic Acid Sequence-based Amplificationmentioning

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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“…Following laser ablation, the PMMA was sonicated in 15 % isopropanol for 5 min (Branson Ultrasonic Corp., USA). The channel was bonded using UV/ozone-assisted thermal bonding (Nugen et al 2009;Truckenmuller et al 2004). This treatment has also previously been show to result in a hydrophilic surface and allow for lower temperature thermal bonding.…”

Section: Laser Ablation and Surface Modificationmentioning

confidence: 99%

“…Recently, the use of polymethyl methacrylate (PMMA) has become a prevalent substrate for use in microfluidics (Nugen et al 2009;Reedy et al 2011;Wang et al 2003), due to its low cost, chemical inertness and manufacturability.…”

Section: Introductionmentioning

confidence: 99%

Development and characterization of a capillary-flow microfluidic device for nucleic acid detection

Jin

Dai

et al. 2012

Microsyst Technol

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We have developed a capillary flow-driven microfluidic biosensor to meet the needs of diagnostics for resource-limited areas. The device combined elements of lateral flow assays and microfluidic technology resulting in a hybrid with benefits of both formats. The biosensor was achieved by bonding two pieces of polymethyl methacrylate with channels ablated by a CO 2 laser, and enclosing an absorbent pad. The channels were UV/ozone treated to increase hydrophilicity which enabled capillary flow. The absorbent pad allowed for continuous flow in the channels once filled. The application of biosensor was demonstrated by detection of DNA with a sandwich assay. The target DNA was hybridized with nucleic acid modified magnetic beads as well as Ru(bpy) 3 2? doped silica nanoparticles. Fluorescent signals were quantified in a holder fabricated to fit in a fluorescent microtiter plate reader. The capillary flow microfluidic was capable to detect 1 pmol target. The assay format which features rapid analysis and does not require the use of pumps could allow for inexpensive point of care diagnostics in the future.

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PMMA biosensor for nucleic acids with integrated mixer and electrochemical detection

Cited by 86 publications

References 39 publications

Miniaturized Detection Systems

Miniaturized Detection Systems

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

Development and characterization of a capillary-flow microfluidic device for nucleic acid detection

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