Shunting microfluidic PCR device for rapid bacterial detection

Research output: Contribution to journalArticle

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Abstract

Polymerase chain reaction (PCR) is commonly used for the analysis of nucleic acids in a variety of applications including clinical. There is, however, a need for a low cost portable PCR device that allows rapid identification of pathogenic bacteria. We report a shunting PCR microfluidic device comprising: polycarbonate microfluidic PCR chip; shunting thermal cycler and fluorescence detector. The microfluidic PCR chip – fabricated using micro-milling and thermal fusion bonding for sealing of the cover – was shunted between three double side temperature zones for thermal cycling. Rapid amplification was observed with heating and cooling rates of 1.8 °C/s and 2 °C/s respectively. Lock-in photodetector for fluorescence detection of the microfluidic PCR chip achieved at 95% confidence an LOD of 75pM FITC and 0.7 ng μl−1 of dsDNA using a QuantiFluor assay kit. The device was validated using universal primers - based on chromosomal DNA extracted from non-pathogenic K-12 subtype of Escherichia coli (E. coli) – for amplification of fragments of 250, 552 and 1500 bp. PCR amplification was demonstrated, with annealing temperatures ranging between 54 °C and 68 °C, and confirmed using gel electrophoresis. The developed shunting PCR microfluidic device will allow for low cost and portable nucleic acid amplification for the detection of infectious diseases.
Original languageEnglish
Article number120303
Number of pages8
JournalTalanta
Volume207
Early online date28 Aug 2019
DOIs
Publication statusPublished - 15 Jan 2020

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Polymerase chain reaction
Microfluidics
Amplification
polycarbonate
Nucleic Acids
Fluorescence
Fluorescein-5-isothiocyanate
Thermal cycling
Photodetectors
Electrophoresis
Escherichia coli
Costs
Assays
Bacteria
Fusion reactions
Gels
Annealing
Cooling
Detectors
Heating

Cite this

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title = "Shunting microfluidic PCR device for rapid bacterial detection",
abstract = "Polymerase chain reaction (PCR) is commonly used for the analysis of nucleic acids in a variety of applications including clinical. There is, however, a need for a low cost portable PCR device that allows rapid identification of pathogenic bacteria. We report a shunting PCR microfluidic device comprising: polycarbonate microfluidic PCR chip; shunting thermal cycler and fluorescence detector. The microfluidic PCR chip – fabricated using micro-milling and thermal fusion bonding for sealing of the cover – was shunted between three double side temperature zones for thermal cycling. Rapid amplification was observed with heating and cooling rates of 1.8 °C/s and 2 °C/s respectively. Lock-in photodetector for fluorescence detection of the microfluidic PCR chip achieved at 95{\%} confidence an LOD of 75pM FITC and 0.7 ng μl−1 of dsDNA using a QuantiFluor assay kit. The device was validated using universal primers - based on chromosomal DNA extracted from non-pathogenic K-12 subtype of Escherichia coli (E. coli) – for amplification of fragments of 250, 552 and 1500 bp. PCR amplification was demonstrated, with annealing temperatures ranging between 54 °C and 68 °C, and confirmed using gel electrophoresis. The developed shunting PCR microfluidic device will allow for low cost and portable nucleic acid amplification for the detection of infectious diseases.",
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Shunting microfluidic PCR device for rapid bacterial detection. / Salman, Abbas; Carney, Helen; Bateson, Simon; Ali, Zulfiqur.

In: Talanta, Vol. 207, 120303, 15.01.2020.

Research output: Contribution to journalArticle

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