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  4. Continuous-flow, microfluidic, qRT-PCR system for RNA virus detection
 
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2018
Zeitschriftenaufsatz
Titel

Continuous-flow, microfluidic, qRT-PCR system for RNA virus detection

Abstract
One of the main challenges in the diagnosis of infectious diseases is the need for rapid and accurate detection of the causative pathogen in any setting. Rapid diagnosis is key to avoiding the spread of the disease, to allow proper clinical decisions to be made in terms of patient treatment, and to mitigate the rise of drug-resistant pathogens. In the last decade, significant interest has been devoted to the development of point-of-care reverse transcription polymerase chain reaction (PCR) platforms for the detection of RNA-based viral pathogens. We present the development of a microfluidic, real-time, fluorescence-based, continuous-flow reverse transcription PCR system. The system incorporates a disposable microfluidic chip designed to be produced industrially with cost-effective roll-to-roll embossing methods. The chip has a long microfluidic channel that directs the PCR solution through areas heated to different temperatures. The solution first travels through a reverse transcription zone where RNA is converted to complementary DNA, which is later amplified and detected in real time as it travels through the thermal cycling area. As a proof of concept, the system was tested for Ebola virus detection. Two different master mixes were tested, and the limit of detection of the system was determined, as was the maximum speed at which amplification occurred. Our results and the versatility of our system suggest its promise for the detection of other RNA-based viruses such as Zika virus or chikungunya virus, which constitute global health threats worldwide.
Author(s)
Fernández-Carballo, B. Leticia
Fraunhofer USA - Center for Manufacturing Innovation; Grup d'Enginyeria de Materials (GEMAT)
McBeth, Christine
Fraunhofer USA - Center for Manufacturing Innovation
McGuiness, Ian
Fraunhofer USA - Center for Manufacturing Innovation
Kalashnikov, Maxim
Fraunhofer USA - Center for Manufacturing Innovation
Baum, Christoph
Fraunhofer-Institut für Produktionstechnologie IPT
Borrós, Salvador
Grup d'Enginyeria de Materials (GEMAT)
Sharon, Andre
Fraunhofer USA - Center for Manufacturing Innovation; Boston University
Sauer-Budge, Alexis F.
Fraunhofer USA - Center for Manufacturing Innovation; Boston University
Zeitschrift
Analytical and bioanalytical chemistry
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DOI
10.1007/s00216-017-0689-8
Language
Englisch
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IPT
Tags
  • LAB on a Chip

  • quantitative reverse ...

  • RNA-based virus detec...

  • infectious discases

  • point of care

  • ebola virus

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