A self-heating cartridge for molecular diagnostics

Liu, Changchun; Mauk, Michael G.; Hart, Robert W.; Qiu, Xianbo; Bau, Haim H.

doi:10.1039/c1lc20345b

Cited by 87 publications

(111 citation statements)

References 44 publications

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Section: Strategies To Miniaturize Isothermal Amplification Reactionsmentioning

confidence: 99%

“…Thus, many microsystems that combined LAMP with different detection methods, newly designed lab-on-chips, microfluidic cartridges and automated techniques were reported. In many studies, the LAMP amplification signals were already detected after 15-40 minutes after the thermal reaction started [50][51][52][53][54].The preferred and the most sensitive method for detection is based on fluorescence (with SYBR green, SYTO green or EvaGreen as fluorescent dyes) that can be detected as early as 15 minutes after the thermal reaction starts. For calcein, a limit of detection (LOD) of about 270 copies/µL was described [55].…”

mentioning

confidence: 99%

“…The HBV template could be detected by an SPR-LAMP system in 17 minutes even at the detection limit of 2 fg/mL. Several LAMP systems of on-chip detection modules were also combined with heating modules [52][53][54]. For the purpose of complete automatic control of the assay, Wang et al integrated sample pretreatment into the microfluidic LAMP device by using magnetic beads and a specially designed isolation membrane [53,54].…”

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confidence: 99%

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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: Strategies To Miniaturize Isothermal Amplification Reactionsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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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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“…40 Unlike PCR, LAMP does not require thermal cycling [41][42][43] , allowing a simpler and cheaper, disposable heating system to be used , [44][45][46][47][48] , and it is particularly temperature robust 40,48,49 . We 80 observed the LAMP reaction to be stable at temperatures of 65±5°C (T able S-6).…”

mentioning

confidence: 99%

A versatile-deployable bacterial detection system for food and environmental safety based on LabTube-automated DNA purification, LabReader-integrated amplification, readout and analysis

Hoehl

Bocholt

Kloke³

et al. 2014

Analyst

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Contamination of foods is a public health hazard that episodically causes thousands of deaths and sickens millions worldwide. T o ensure food safety and quality, rapid, low-cost and easy-to-use detection methods are desirable. Here, the LabSystem is introduced for integrated, automated DNA purification, 10 amplification and detection. It consists of a disposable, centrifugally -driven DNA purification platform (LabT ube) and the subsequent amplification and detection in a low-cost UV/vis-reader (LabReader). For demonstration of the LabSystem in the context of food safety, purification of Escherichia coli (nonpathogenic E. coli and pathogenic verotoxin-producing E. coli (VT EC)) in water and milk, and the product-spoiler Alicyclobacillus acidoterrestris (A. acidoterrestris) in apple juice was integrated and 15 optimized in the LabT ube. Inside the LabReader, the purified DNA was amplified, readout and analyzed using both qualitative isothermal loop-mediated DNA amplification (LAMP) and quantitative real-time PCR. For the LAMP -LabSystem, the combined detection limits for purification and amplification of externally lysed VT EC and A. acidoterrestris is 10 2 -10 3 cell-equivalents. In the PCR-LabSystem for E. coli cells, the quantification limit is 10 2 cell-equivalents including LabT ube-integrated lysis. T he 20 demonstrated LabSystem only requires a laboratory centrifuge (to operate the disposable, fully closed LabT ube) and the low-cost LabReader for DNA amplification, readout and analysis. Compared with commercial DNA amplification devices, the LabReader improves sensitivity and specificity by the simultaneous readout of four wavelengths and the continuous readout during temperature cycling. T he use of a detachable eluate tube as an interface affords semi-automation of the LabSystem, which does not 25 require specialized training. It reduces hands-on time from about 50 to 3 min with only two handling steps: sample input and transfer of the detachable detection tube.

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“…Infrared cameras can readily assess the uniformity of heated surfaces and spatial temperature variation in microfluidic chips. For example, Figure 5 shows a top view of a stage heated by chemical heating [15]. Similar evaluations of Peltier and resistive heating elements.…”

Section: Chip Fabrication Instrumentation and Operationmentioning

confidence: 99%

Microscale Implementation and Image Analysis of Fluid Processes

Mauk¹,

Chiou²,

Varapula³

2016 ASEE Annual Conference &Amp; Exposition Proceedings

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Microscale fluidics offers new avenues for teaching CAD, prototyping, fluid mechanics, heat and mass transfer, process engineering, control, and image analysis. Miniaturized fluid systems are implemented in credit card-sized, clear plastic 'chips' that host a network of conduits, chambers, filters, manifolds, and flow control devices. The chips are connected to programmable syringe pumps. With colored and fluorescent dyes, images and videos of flow characteristics and mixing can be captured, processed, and analyzed with low-cost CCD cameras and along with image processing software (ImageJ or MATLAB®). Moreover, heats of mixing, heats of reaction, and convective, conductive, and radiative heat transfer in fluid systems can be analyzed using thermal image infrared (8-12 microns) cameras. Student projects include CAD of microfluidic chips, fabrication of chips using a CO2 laser cutter, 3D-printer, or CNC mill; experimental setup on a desktop with programmable syringe pumps, sensors, and CCD and thermal infrared camera, experimentation, and analysis of images. We study concurrent and countercurrent heat exchangers, various active and passive mixers, heats of mixing between alcohols and aqueous salt solutions, and acid-base neutralization reactions. These laboratory projects provide instructive and accessible hands-on experimentation, at levels ranging from intuitive and visual to more analytical treatments, in subject areas of fluid mechanics, heat transfer, reaction engineering, image processing and machine vision, engineering modeling, and rapid prototyping. We emphasize skills and concepts gained for their relevancy to energy efficiency, sustainability, and green manufacturing.

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A self-heating cartridge for molecular diagnostics

Cited by 87 publications

References 44 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

A versatile-deployable bacterial detection system for food and environmental safety based on LabTube-automated DNA purification, LabReader-integrated amplification, readout and analysis

Microscale Implementation and Image Analysis of Fluid Processes

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