Centrifugal LabTube platform for fully automated DNA purification and LAMP amplification based on an integrated, low-cost heating system

Hoehl, Melanie M.; Weißert, Michael; Dannenberg, Arne; Nesch, Thomas; Paust, Nils; Stetten, Felix von; Zengerle, Roland; Slocum, Alexander H.; Steigert, J.

doi:10.1007/s10544-014-9841-9

Cited by 12 publications

(11 citation statements)

References 49 publications

(51 reference statements)

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“…Many efforts have been made to solve this problem. For example, LabTube system with the help of laboratory centrifuges ( Figure 12 A) [ 152 , 153 ] and lab on DVDs system using DVD discs and commercial drives [ 96 , 126 ] have been suggested to make centrifugal microfluidic devices more universal and portable. In NA analysis, simple and compact colorimetric detection is used instead of fluorescence detection in some cases as a result of the development of isothermal amplification methods [ 89 , 91 ].…”

Section: Discussionmentioning

confidence: 99%

A Review of Biomedical Centrifugal Microfluidic Platforms

et al. 2016

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Centrifugal microfluidic or lab-on-a-disc platforms have many advantages over other microfluidic systems. These advantages include a minimal amount of instrumentation, the efficient removal of any disturbing bubbles or residual volumes, and inherently available density-based sample transportation and separation. Centrifugal microfluidic devices applied to biomedical analysis and point-of-care diagnostics have been extensively promoted recently. This paper presents an up-to-date overview of these devices. The development of biomedical centrifugal microfluidic platforms essentially covers two categories: (i) unit operations that perform specific functionalities, and (ii) systems that aim to address certain biomedical applications. With the aim to provide a comprehensive representation of current development in this field, this review summarizes progress in both categories. The advanced unit operations implemented for biological processing include mixing, valving, switching, metering and sequential loading. Depending on the type of sample to be used in the system, biomedical applications are classified into four groups: nucleic acid analysis, blood analysis, immunoassays, and other biomedical applications. Our overview of advanced unit operations also includes the basic concepts and mechanisms involved in centrifugal microfluidics, while on the other hand an outline on reported applications clarifies how an assembly of unit operations enables efficient implementation of various types of complex assays. Lastly, challenges and potential for future development of biomedical centrifugal microfluidic devices are discussed.

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Section: Discussionmentioning

confidence: 99%

A Review of Biomedical Centrifugal Microfluidic Platforms

et al. 2016

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“…Thus, a variety of biochemical applications on the centrifugal microdevice platform including cell lysis and PCR (Kellogg et al, 2000), ion sensor (Grumann et al, 2005), alcohol assay (Steigert et al, 2006), and immunoassay have been reported (Haeberle et al, 2006). Hoehl et al (2014) showed a centrifugal LabTube platform to automate DNA purification and LAMP amplification to detect verotoxin-producing Escherichia coli (VTEC). As for the pathogen detection applications, Lutz et al (2011) presented a centrifugal lab-ona-foil platform for RPA based Staphylococcus aureus identification.…”

Section: Introductionmentioning

confidence: 99%

Integrated centrifugal reverse transcriptase loop-mediated isothermal amplification microdevice for influenza A virus detection

Jung

Park

et al. 2015

Biosensors and Bioelectronics

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An integrated reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) microdevice which consists of microbead-assisted RNA purification and RT-LAMP with real-time monitoring by a miniaturized optical detector was demonstrated. The integrated RT-LAMP microdevice includes four reservoirs for a viral RNA sample (purified influenza A viral RNA or lysates), a washing solution (70% ethanol), an elution solution (RNase-free water), and an RT-LAMP cocktail, and two chambers (a waste chamber and an RT-LAMP reaction chamber). The separate reservoirs for a washing solution, an elution solution, and an RT-LAMP cocktail were designed with capillary valves for stable storage. Three influenza A virus strains (A/H1N1, A/H3N2, and A/H5N1) were used for RNA templates, and RT-LAMP primer sets were designed to detect hemagglutinin (HA) and conserved M gene. Sequential sample flow to the microbeads for RNA purification was achieved by centrifugal force with optimization of capillary valves and a siphon channel. Furthermore, the purified RNA solution was successfully isolated from the waste solution by changing the rotational direction, and combined with the RT-LAMP cocktail in the RT-LAMP reaction chamber for target gene amplification. Total process from the sample injection to the result was completed in 47 min. Influenza A H1N1 virus was confirmed on the integrated RT-LAMP microdevice even with 10 copies of viral RNAs, which revealed 10-fold higher sensitivity than that of a conventional RT-PCR. Subtyping and specificity test of influenza A H1N1 viral lysates were also performed and clinical samples were successfully genotyped to confirm influenza A virus on our proposed integrated microdevice.

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“…Microfluidic chips/cards with the capacity to target multiple infectious diseases in parallel are a promising means to rapidly diagnose and satisfy many POC criteria (McCalla and Tripathi 2011 ). Innovative strategies for precise microfluidic control or loading and sealing samples into multiple well compartments (for parallel detection) have been described and reviewed (Jung et al 2015 ; Sackmann et al 2014 ; Su et al 2014 ) including: the manual deposition of sample in a small number of individual wells (Fang et al 2010 ; Lee et al 2008 ), the utilization of peripheral equipment to load samples with arrayers and injectors (Matsubara et al 2004 ; Morrison et al 2006 ; Duarte et al 2013 ), pumps (Furuberg et al 2008 ; Sun et al 2015 ), centrifuges (Lutz et al 2010 ; Jung et al 2014 ; Hoehl et al 2014 ), development of multiple-layer cards utilizing slip (Shen et al 2011 ), air, thermal, magnets, physical valve control (Fang et al 2012 ; Stedtfeld et al 2012 ; Trung et al 2010 ; Wang et al 2011 ), vacuum assistance (Abe et al 2011 ), or immiscible fluids to prime cards (Gansen et al 2012 ), surface treatment for primer immobilization and mobilization during filling (Morrison et al 2006 ), and finally, propagation of sample through a parallel network of interconnecting channels using an air vent for each reaction well (Fang et al 2010 ; Ramalingam et al 2010 ; Stedtfeld et al 2012 ; Tourlousse et al 2012 ). However, most microfluidic cards made of glass, silicon, and polymers require highly sophisticated microfabrication (e.g.…”

Section: Introductionmentioning

confidence: 99%

Static self-directed sample dispensing into a series of reaction wells on a microfluidic card for parallel genetic detection of microbial pathogens

Stedtfeld

Liu

Stedtfeld

et al. 2015

Biomed Microdevices

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A microfluidic card is described for simultaneous and rapid genetic detection of multiple microbial pathogens. The hydrophobic surface of native acrylic and a novel microfluidic mechanism termed “airlock” were used to dispense sample into a series of 64 reaction wells without the use of valves, external pumping peripherals, multiple layers, or vacuum assistance. This airlock mechanism was tested with dilutions of whole human blood, saliva, and urine, along with mock samples of varying viscosities and surface tensions. Samples spiked with genomic DNA (gDNA) or crude lysates from clinical bacterial isolates were tested with loop mediated isothermal amplification assays (LAMP) designed to target virulence and antibiotic resistance genes. Reactions were monitored in real time using the Gene-Z, which is a portable smartphone-driven system. Samples loaded correctly into the microfluidic card in 99.3 % of instances. Amplification results confirmed no carryover of pre-dispensed primer between wells during sample loading, and no observable diffusion between adjacent wells during the 60 to 90 min isothermal reaction. Sensitivity was comparable between LAMP reactions tested within the microfluidic card and in conventional vials. Tests demonstrate that the airlock card works with various sample types, manufacturing techniques, and can potentially be used in many point-of-care diagnostics applications.Electronic supplementary materialThe online version of this article (doi:10.1007/s10544-015-9994-1) contains supplementary material, which is available to authorized users.

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Centrifugal LabTube platform for fully automated DNA purification and LAMP amplification based on an integrated, low-cost heating system

Cited by 12 publications

References 49 publications

A Review of Biomedical Centrifugal Microfluidic Platforms

A Review of Biomedical Centrifugal Microfluidic Platforms

Integrated centrifugal reverse transcriptase loop-mediated isothermal amplification microdevice for influenza A virus detection

Static self-directed sample dispensing into a series of reaction wells on a microfluidic card for parallel genetic detection of microbial pathogens

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