A real-time convective PCR machine in a capillary tube instrumented with a CCD-based fluorometer
Journal
Sensors and Actuators, B: Chemical
Journal Volume
183
Pages
434-440
Date Issued
2013
Author(s)
Abstract
This research reports the design, analysis, integration, and test of a prototype of a real-time convective polymerase chain reaction (RT-cPCR) machine that uses a color charged coupled device (CCD) for detecting the emission of fluorescence intensity from an RT-cPCR mix in a microliter volume glass capillary. Because of its simple mechanism, DNA amplification involves employing the cPCR technique with no need for thermocycling control. The flow pattern and temperature distribution can greatly affect the cPCR process in the capillary tube, a computational fluid dynamics (CFD) simulation was conducted in this study for the first time to estimate the required period of an RT-cPCR cycle. This study also tested the PCR mix containing hepatitis B virus (HBV) plasmid samples by using SYBR Green I fluorescence labeling dye to assess the prototype performance. The measured results from the image-processing scheme indicate that the RT-cPCR prototype with a CCD-based fluorometer can achieve similar DNA quantification reproducibility compared to commercial machines, even when the initial DNA concentration in the test PCR mix is reduced to 10 copies/μL ? 2013 Elsevier B.V.
SDGs
Other Subjects
Capillary tubes; Charge coupled devices; Computational fluid dynamics; DNA; Fluorescence; Fluorometers; Tubes (components); Viruses; Computational fluid dynamics simulations; cPCR; DNA quantification; Fluorescence intensities; Fluorescence labeling; Hepatitis B virus; Reproducibilities; RT-cPCR; Polymerase chain reaction
Publisher
Elsevier
Type
journal article