Recent outbreaks of deadly infectious diseases, such as Ebola and Middle East respiratory syndrome coronavirus, have motivated the research for accurate, rapid diagnostics that can be administered at the point of care. Nucleic acid biomarkers for these diseases can be amplified and quantified via polymerase chain reaction (PCR). In order to solve the problems of conventional PCR-speed, uniform heating and cooling, and massive metal heating blocks-an innovative optofluidic cavity PCR method using light-emitting diodes (LEDs) is accomplished. Using this device, 30 thermal cycles between 94 °C and 68 °C can be accomplished in 4 min for 1.3 μL (10 min for 10 μL). Simulation results show that temperature differences across the 750 μm thick cavity are less than 2 °C and 0.2 °C, respectively, at 94 °C and 68 °C. Nucleic acid concentrations as low as 10-8 ng μL-1 (2 DNA copies per μL) can be amplified with 40 PCR thermal cycles. This simple, ultrafast, precise, robust, and low-cost optofluidic cavity PCR is favorable for advanced molecular diagnostics and precision medicine. It is especially important for the development of lightweight, point-of-care devices for use in both developing and developed countries. Novel optical cavity polymerase chain reaction (PCR) driven by a light-emitting diode (LED) is demonstrated for fast, accurate, and reliable PCR-based diagnostics. Using optical cavity PCR, nucleic acid amplification with the template DNA concentration has been successfully demonstrated, ranging from 10-5 to 10-8 ng μL-1 within 5-15 min with excellent repeatability and reproducibility.
Bibliographical noteFunding Information:
This work was supported in part by a grant from the PCARI-IHITM (Grant No. 131) and in part by the Global Research Lab Program (2013- 050616) and in part by the Air Force Office of Scientific Research Grants AFOSR FA2386-13-1-4120. We thank Byungrae Cho for help with heat transfer simulation and Charley Y. Cui for help with fabrication of the optical cavity PCR chamber.
© 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
- Light-emitting diodes
- Molecular diagnostics
- Optical cavities
- Point-of-care diagnostics
- Polymerase chain reaction