Journal
BIOMICROFLUIDICS
Volume 9, Issue 4, Pages -Publisher
AMER INST PHYSICS
DOI: 10.1063/1.4928486
Keywords
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Categories
Funding
- NIH BRAIN Initiative Grant (NEI)
- NIH BRAIN Initiative Grant [NIMH 1-U01-MH106027-01]
- NIH Single Cell Grant [1 R01 EY023173]
- NSF [EHR 0965945, CISE 1110947]
- NIH Computational Neuroscience Training grant [DA032466-02]
- Georgia Tech Translational Research Institute for Biomedical Engineering AMP
- Science (TRIBES) Seed Grant Awards Program
- Georgia Tech Fund for Innovation in Research and Education (GT-FIRE)
- Wallace H. Coulter Translational/Clinical Research Grant Program
- Georgia Tech through the Institute for Bioengineering and Biosciences Junior Faculty Award
- Technology Fee Fund
- Invention Studio
- George W. Woodruff School of Mechanical Engineering
- DOE [DE-AC05-06OR23100]
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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 mu l, 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 degrees C and 68 degrees 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. (C) 2015 AIP Publishing LLC.
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