Extraction, amplification and detection of DNA in microfluidic chip-based assays

Wu, Jinbo; Kodzius, Rimantas; Cao, Wenbin; Wen, Weijia

doi:10.1007/s00604-013-1140-2

Cited by 104 publications

(76 citation statements)

References 244 publications

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“…Fabrications of polymeric microfluidics encountered with some problems such as preparation of the polymer molding solution, multilayer polymers, and penetration of the reaction mixtures in a polymer-based platform. Furthermore, fabrications of the microfluidic containers often were involved some surface treatments with the hydrophobic polymers for preventing the air bubble formations and improving the biocompatibilities, consequently [35,36]; however, these limitations were not encountered in the capillary tubes for microfluidic LAMPs (Table 3).…”

Section: Discussionmentioning

confidence: 99%

Microfluidic method for rapid turbidimetric detection of the DNA of Mycobacterium tuberculosis using loop-mediated isothermal amplification in capillary tubes

Rafati

Gill

2014

Microchim Acta

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We describe a microfluidic method for rapid isothermal turbidimetric detection of the DNA of Mycobacterium tuberculosis. Loop-mediated isothermal amplification is accomplished in capillary tubes for amplifying DNA in less than 15 min, and sensitivity and specificity were compared to conventional loop-mediated isothermal amplification (LAMP). The method can detect as little as 1 pg mL −1 DNA in a sample. Results obtained with clinical specimens indicated 90 % sensitivity and 95 % specificity for microfluidic LAMP in comparison to culture methods. No interference occurred due to the presence of nonspecific DNAs. The findings demonstrate the power of the new microfluidic LAMP test for rapid molecular detection of microorganisms even when using bare eyes.

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Section: Discussionmentioning

confidence: 99%

Microfluidic method for rapid turbidimetric detection of the DNA of Mycobacterium tuberculosis using loop-mediated isothermal amplification in capillary tubes

Rafati

Gill

2014

Microchim Acta

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“…Microfluidic devices will significantly influence the future of the biological process industry 2 . Amongst these, microfluidic mixers which allow the rapid mixing of fluids within the microchannel find application in many biochemical processes, including the synthesis or sequencing of nucleic acids, 3 chemical reactions, 4 DNA purification, 5 and polymerase chain reaction 6 . A further advantage of integrating microfluidic mixers is a reduction in the incubation time for bead-based enzyme-linked immunosorbent assays (ELISA) used in the study of cancer biomarkers 7 …”

Section: Introductionmentioning

confidence: 99%

A “twisted” microfluidic mixer suitable for a wide range of flow rate applications

et al. 2016

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This paper proposes a new “twisted” 3D microfluidic mixer fabricated by a laser writing/microfabrication technique. Effective and efficient mixing using the twisted micromixers can be obtained by combining two general chaotic mixing mechanisms: splitting/recombining and chaotic advection. The lamination of mixer units provides the splitting and recombination mechanism when the quadrant of circles is arranged in a two-layered serial arrangement of mixing units. The overall 3D path of the microchannel introduces the advection. An experimental investigation using chemical solutions revealed that these novel 3D passive microfluidic mixers were stable and could be operated at a wide range of flow rates. This micromixer finds application in the manipulation of tiny volumes of liquids that are crucial in diagnostics. The mixing performance was evaluated by dye visualization, and using a pH test that determined the chemical reaction of the solutions. A comparison of the tornado-mixer with this twisted micromixer was made to evaluate the efficiency of mixing. The efficiency of mixing was calculated within the channel by acquiring intensities using ImageJ software. Results suggested that efficient mixing can be obtained when more than 3 units were consecutively placed. The geometry of the device, which has a length of 30 mm, enables the device to be integrated with micro total analysis systems and other lab-on-chip devices.

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“…Moreover, the nested PCR assay is quite laborious and time-consuming and requires gel electrophoresis for amplicon detection, making it prone to cross contamination. Recently developed miniaturized chips have progressed by the introduction of microfluid technology for pathogen detection [49][50][51]. However, high costs of the reaction vessels and instruments, problems in product manufacturing, and sub-optimal detection methods have deferred these methods from being applied to point-ofcare diagnostics for resource-poor facilities.…”

Section: Discussionmentioning

confidence: 99%

Rapid and sensitive detection of canine distemper virus by one-tube reverse transcription-insulated isothermal polymerase chain reaction

et al. 2014

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Background: Canine distemper virus (CDV) has been associated with outbreaks of canine infectious respiratory disease in shelters and boarding kennel environments. POCKIT TM Nucleic Acid Analyzer is a field-deployable device capable of generating automatically interpreted insulated isothermal polymerase chain reaction (iiPCR) results from extracted nucleic acid within one hour. In this study, reverse transcription iiPCR (RT-iiPCR) was developed to facilitate point-of-need diagnosis of CDV infection. Results: Analytical sensitivity (limit of detection 95%) of the established CDV RT-iiPCR was about 11 copies of in vitro transcribed RNA per reaction. CDV RT-iiPCR generated positive signals from CDV, but not Bordetella bronchiseptica, canine parvovirus, canine herpesvirus, canine adenovirus 2, canine influenza virus (subtype H3N8), canine parainfluenza virus, and canine respiratory coronavirus. To evaluate accuracy of the established reaction in canine distemper clinical diagnosis, 110 specimens from dogs, raccoons, and foxes suspected with CDV infection were tested simultaneously by CDV RT-iiPCR and real-time RT-PCR. CDV RT-iiPCR demonstrated excellent sensitivity (100%) and specificity (100%), compared to real-time RT-PCR.

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Extraction, amplification and detection of DNA in microfluidic chip-based assays

Cited by 104 publications

References 244 publications

Microfluidic method for rapid turbidimetric detection of the DNA of Mycobacterium tuberculosis using loop-mediated isothermal amplification in capillary tubes

Microfluidic method for rapid turbidimetric detection of the DNA of Mycobacterium tuberculosis using loop-mediated isothermal amplification in capillary tubes

A “twisted” microfluidic mixer suitable for a wide range of flow rate applications

Rapid and sensitive detection of canine distemper virus by one-tube reverse transcription-insulated isothermal polymerase chain reaction

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