Sub-5-Minute Ultrafast PCR using Digital Microfluidics

Liang Wan; Mingzhong Li; Man-Kay Law; Pui-In Mak; Rui P Martins; Yanwei Jia

doi:10.1016/j.bios.2023.115711

Sub-5-Minute Ultrafast PCR using Digital Microfluidics

Biosens Bioelectron. 2023 Dec 15:242:115711. doi: 10.1016/j.bios.2023.115711. Epub 2023 Sep 26.

Authors

Liang Wan¹, Mingzhong Li², Man-Kay Law³, Pui-In Mak³, Rui P Martins⁴, Yanwei Jia⁵

Affiliations

¹ The State Key Laboratory of Analog and Mixed-Signal VLSI, Institute of Microelectronics, University of Macau, Macao, China.
² The State Key Laboratory of Analog and Mixed-Signal VLSI, Institute of Microelectronics, University of Macau, Macao, China; Silergy Semiconductor (Macau) Limited, Macao, China.
³ The State Key Laboratory of Analog and Mixed-Signal VLSI, Institute of Microelectronics, University of Macau, Macao, China; Faculty of Science and Technology - Electrical and Computer Engineering, University of Macau, Macao, China.
⁴ The State Key Laboratory of Analog and Mixed-Signal VLSI, Institute of Microelectronics, University of Macau, Macao, China; Faculty of Science and Technology - Electrical and Computer Engineering, University of Macau, Macao, China; On Leave from Instituto Superior Técnico, Universidade de Lisboa, Lisboa, Portugal.
⁵ The State Key Laboratory of Analog and Mixed-Signal VLSI, Institute of Microelectronics, University of Macau, Macao, China; Faculty of Science and Technology - Electrical and Computer Engineering, University of Macau, Macao, China; MoE Frontiers Science Center for Precision Oncology, University of Macau, Macao, China. Electronic address: yanweijia@um.edu.mo.

PMID: 37797533
DOI: 10.1016/j.bios.2023.115711

Abstract

The development of a rapid and reliable polymerase chain reaction (PCR) method for point-of-care (POC) diagnosis is crucial for the timely identification of pathogens. Microfluidics, which involves the manipulation of small volumes of fluidic samples, has been shown to be an ideal approach for POC analysis. Among the various microfluidic platforms available, digital microfluidics (DMF) offers high degree of configurability in manipulating μL/nL-scale liquid and achieving automation. However, the successful implementation of ultrafast PCR on DMF platforms presents challenges due to inherent system instability. In this study, we developed a robust and ultrafast PCR in 3.7-5 min with a detection sensitivity comparable to conventional PCR. Specifically, the implementation of the pincer heating scheme homogenises the temperature within a drop. The utilization of a μm-scale porous hydrophobic membrane suppresses the formation of bubbles under high temperatures. The design of a groove around the high-temperature zone effectively mitigates the temperature interference. The integration of a soluble sensor into the droplets provides an accurate and instant in-drop temperature sensing. We envision that the fast, robust, sensitive, and automatic DMF system will empower the POC testing for infectious diseases.

Keywords: Chemical temperature sensor; Digital microfluidics; Electrowetting-on-dielectrics; Ultrafast polymerase chain reaction.

MeSH terms

Biosensing Techniques*
Communicable Diseases*
Humans
Microfluidics / methods
Point-of-Care Systems
Polymerase Chain Reaction