SU-8 based continuous-flow RT-PCR bio-chips under high-precision temperature control

Tsai, Nan-Chyuan; Sue, Chung Yang

doi:10.1016/j.bios.2005.12.023

Cited by 33 publications

(21 citation statements)

References 8 publications

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“…This design advantage is achieved by maintaining the zones at preset constant temperatures . Currently, the structural styles of the continuousflow RT-PCR microfluidics can be divided into two main categories: the serpentine-channel continuous-flow RT-PCR (Felbel et al 2008;Tsai and Sue 2006) and the spiralchannel continuous-flow RT-PCR (Hartung et al 2009). In the latter configuration, PCR device has an underlying advantage of a circular arrangement of temperature zones in the sequence of denaturation, annealing, and extension, which could increase the PCR efficiency compared with the former.…”

Section: Introductionmentioning

confidence: 99%

Fast identification of foodborne pathogenic viruses using continuous-flow reverse transcription-PCR with fluorescence detection

Zhang

Xing

2010

Microfluid Nanofluid

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Being beneficial from dramatic process in biochemical and micro-electro-mechanical technologies, this study presents an integrated microfluidic system capable of performing a continuous-flow reverse transcription-polymerase chain reaction (RT-PCR) combined with fluorescence microscopy for rapid diagnosis of RNA-based viruses. The device consists of two heated cylinders for heating the different reaction zones of the reverse transcription and the amplification. In the amplification cylinder, bath refrigerating is used for thermal protection of the annealing region, which is heated possibly due to the thermal effects of radiation, conduction, and/or convection from denaturation and extension regions, thus resulting in a space-saving design of the whole device. Detection of the amplified products is performed on-line by a fluorescence microscopy with SybrGreen I, a widely used intercalating dye. In this article, the proposed miniature RT-PCR system is used to amplify and detect two RNA-based viruses (Noroviruses (NVs) and Rotaviruses (RVs)), which are now recognized as the most common etiological agents of acute viral gastroenteritis causing numerous outbreaks worldwide. The experimental data have demonstrated the ability of the presented system to perform a two-step or one-step RT-PCR process. On this device, the NVs and RVs RNA samples were successfully reverse transcribed and amplified within 1 h, and the limit of detection of the RNA concentration was 6.4 9 10 4 copies ll -1 using onestep RT-PCR process. Consequently, the developed microfluidic system can provide a promising platform for fast diagnosis of RNA-based viruses.

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Section: Introductionmentioning

confidence: 99%

Fast identification of foodborne pathogenic viruses using continuous-flow reverse transcription-PCR with fluorescence detection

Zhang

Xing

2010

Microfluid Nanofluid

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“…The PCR amplification process contains three sectors with different process temperatures: denaturation, annealing, and extension. The annealing sector, which contains the attachment of separated single strand DNA to a specific primer, takes place at 54˚C [65]. Subsequently, the design process for appropriate microfluidic chip in this application is presented.…”

Section: Steady State Modelmentioning

confidence: 99%

Geometric optimization for a thermal microfluidic chip

Rahmat

Hubert

2010

J Mech Sci Technol

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During the past two decades, thermal microfluidic chips have significantly been investigated. Due to their high capacity of heat transport, an increasing number of studies on different aspects of thermal microfluidic chips have been conducted.However, a comprehensive investigation on the geometry of microfluidic chips using state of the art finite element software is absent from the literature. In this thesis, geometric parameters of thermal microfluidic chips have been optimized, using finite element software. Consequently, micro and macro phenomena were

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“…A simplified model and a three-dimensional model were used to predict the PCR mixture velocity in and the temperature distribution around the Teflon tubing. Tsai and Sue [18] designed a micro-RT-PCR chip to detect tumor viruses. Commercial software was utilized to simulate the thermal entry length of fluid at different temperature regions.…”

Section: Introductionmentioning

confidence: 99%

Analysis of Thermal Performance in a Bidirectional Thermocycler by Including Thermal Contact Characteristics

Chen

et al. 2014

Micromachines

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This paper illustrates an application of a technique for predicting the thermal characteristics of a bidirectional thermocycling device for polymerase chain reaction (PCR). The micromilling chamber is oscillated by a servo motor and contacted with different isothermal heating blocks to successfully amplify the DNA templates. Because a comprehensive database of contact resistance factors does not exist, it causes researchers to not take thermal contact resistance into consideration at all. We are motivated to accurately determine the thermal characteristics of the reaction chamber with thermal contact effects existing between the heater surface and the chamber surface. Numerical results show that the thermal contact effects between the heating blocks and the reaction chamber dominate the temperature variations and the ramping rates inside the PCR chamber. However, the influences of various temperatures of the ambient conditions on the sample temperature during three PCR steps can be negligible. The experimental temperature profiles are compared well with the numerical simulations by considering the thermal contact conductance coefficient which is empirical by the experimental fitting. To take thermal contact conductance coefficients into consideration in the thermal simulation is recommended to predict a reasonable temperature profile of the reaction chamber during various thermal cycling processes. Finally, the PCR experiments present that Hygromycin B DNA templates are amplified successfully. Furthermore, our group is the first group to OPEN ACCESS Micromachines 2014, 5 1446 introduce the thermal contact effect into theoretical study that has been applied to the design of a PCR device, and to perform the PCR process in a bidirectional thermocycler.

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SU-8 based continuous-flow RT-PCR bio-chips under high-precision temperature control

Cited by 33 publications

References 8 publications

Fast identification of foodborne pathogenic viruses using continuous-flow reverse transcription-PCR with fluorescence detection

Fast identification of foodborne pathogenic viruses using continuous-flow reverse transcription-PCR with fluorescence detection

Geometric optimization for a thermal microfluidic chip

Analysis of Thermal Performance in a Bidirectional Thermocycler by Including Thermal Contact Characteristics

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