Miniature RT-PCR system for diagnosis of RNA-based viruses

Liao, Chia-Sheng; Lee, Gwo‐Bin; Liu, Hsiao Sheng; Hsieh, Tar‐Hwa; Luo, Ching‐Hsing

doi:10.1093/nar/gni157

Cited by 95 publications

(51 citation statements)

References 19 publications

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“…Similar to microfluidic PCR devices, the miniaturized RT-PCR devices can also be divided into two categories: microchamber stationary RT-PCR and continuous-flow RT-PCR. A functional lab-on-a-chip for RT-PCR was reported by Liao et al (2005), where the development of a miniature RT-PCR system was used for the diagnosis of RNA-based viruses. This miniature RT-PCR was performed using a two-step reaction approach (i.e., performing RT and then moving the mixture to another chamber to perform PCR) followed by off-chip detection.…”

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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“…Thus, we can conclude that the relationship between output and input is linear, i.e., we will achieve non-linear calibration of sensor. Finally, we use the IP to obtain a non-linear calibration of concentration sensor in the literature [8]. Practice shows that the use of IP for non-linear calibration has a high calibration accuracy, faster convergence rate and very good stability.…”

Section: Resultsmentioning

confidence: 99%

“…The idea of this system's work is applied in engineering practice, such as the attitude control of satellite; for more details refer to the literature [7]. Diagnosis of RNA-based viruses sometimes needs the help of nonlinear correction [8], so the method must undergo further research to find more benefits for the development of the society.…”

Section: Introductionmentioning

confidence: 99%

Nonlinear Correction of Sensor Based on Immunization Programs

Lu¹,

Niu²,

Zhou³

et al. 2014

TOAUTOCJ

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Sensor applications in medical instrumentation measurement are very extensive, but mostly the input and output of the sensor is nonlinear, which brings a great inconvenience for the measure of medical instruments. This paper introduces biological immune system on the basis of the evolution planning, thereby forming immunization programs to use non-linear calibration of the sensor. This method joins nonlinear correction links after sensor amplification and A/D converter links and uses the immunization program to obtain the polynomial coefficients of the nonlinear correction link. Simulations show that using immunization programs to have non-linear calibration to sensor, gives high correction accuracy, faster convergence speed and very good stability.

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“…14 Similar to commercially available PCR instruments, conductive heating using thermoelectric, or Peltier, elements is common for microfluidic thermocyclers. 10 Resistive 15 and convective 16,17 heating have also been implemented. Radiative heating has been applied to microfluidic PCR with both broadband and focused, monochromatic sources and offers the unique advantage of direct, non-contact heating for greater efficiency and ramping rates as compared with resistive and convective heating instruments.…”

Section: Introductionmentioning

confidence: 99%

“…[21][22][23] The smaller spatial scales and faster heating and cooling rates of microfluidic PCR systems can complicate temperature sensing and control. Techniques used in the previous work include reference chambers with thermocouples, 19 fluorescent dyes, 22 pyrometers, 24 resistance measurements, 15 and open-loop control. 25,26 Most PCR technologies, especially within the field of microfluidics, have advanced the speed and throughput for screening many samples or patients for a single genetic target.…”

Section: Introductionmentioning

confidence: 99%

Thermally multiplexed polymerase chain reaction

et al. 2015

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Amplification of multiple unique genetic targets using the polymerase chain reaction (PCR) is commonly required in molecular biology laboratories. Such reactions are typically performed either serially or by multiplex PCR. Serial reactions are time consuming, and multiplex PCR, while powerful and widely used, can be prone to amplification bias, PCR drift, and primer-primer interactions. We present a new thermocycling method, termed thermal multiplexing, in which a single heat source is uniformly distributed and selectively modulated for independent temperature control of an array of PCR reactions. Thermal multiplexing allows amplification of multiple targets simultaneously-each reaction segregated and performed at optimal conditions. We demonstrate the method using a microfluidic system consisting of an infrared laser thermocycler, a polymer microchip featuring 1 ll, oil-encapsulated reactions, and closed-loop pulsewidth modulation control. Heat transfer modeling is used to characterize thermal performance limitations of the system. We validate the model and perform two reactions simultaneously with widely varying annealing temperatures (48 C and 68 C), demonstrating excellent amplification. In addition, to demonstrate microfluidic infrared PCR using clinical specimens, we successfully amplified and detected both influenza A and B from human nasopharyngeal swabs. Thermal multiplexing is scalable and applicable to challenges such as pathogen detection where patients presenting non-specific symptoms need to be efficiently screened across a viral or bacterial panel. V C 2015 AIP Publishing LLC.

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Miniature RT-PCR system for diagnosis of RNA-based viruses

Cited by 95 publications

References 19 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

Nonlinear Correction of Sensor Based on Immunization Programs

Thermally multiplexed polymerase chain reaction

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