A versatile oscillating‐flow microfluidic PCR system utilizing a thermal gradient for nucleic acid analysis

Abstract

We report the development of a versatile system based on the oscillating‐flow methodology in a thermal gradient system for nucleic acid analysis. Analysis of DNA and RNA samples were performed in the device, without additional temperature control and complexity. The technique reported in this study eliminates the need for predetermined fluidic channels for thermocycles, and complexity involved with additional incubation steps required for RNA amplification. A microfluidic device was fabricated using rapid prototyping by simply sandwiching dual side adhesive Kapton tape and a polydimethylsiloxane spacer between glass microscope slides. Amplification of the 181‐bp segment of a viral phage DNA (ΦX174) and B2M gene in human RNA samples was demonstrated using the system. The developed system enables simultaneous acquisition of amplification and melt curves, eliminating the need for postprocessing. A direct comparison between the oscillating‐flow system and a commercial real‐time polymerase chain reaction (PCR) instrument showed complete agreement in PCR data and improved sample‐to‐result time by eliminating an additional 30 min melt curve step required in commercial PCR systems.

Perspective Chapter: Microfluidic Technologies for On-Site Detection and Quantification of Infectious Diseases – The Experience with SARS-CoV-2/COVID-19

“…The differences in how these techniques can be achieved or improved upon with microfluidics depend on the micro-channel design of the system. In continuous-flow microfluidics, the four main categories of channel design include serpentine, spiral, oscillating-flow, and straight microchannels [22]. Each design channel design serves its own unique purpose when being used in microfluidics.…”

“…microchannels can be used to extend the time in which a fluid is exposed to external factors, such as UV light for crosslinking or heat to promote a particular reaction, across a more unified and even distribution of the fluid [18,22]. Oscillatory continuous-flow microfluidics has been used to increase the effectiveness of liquid-liquid separations as well as aid in sample concentration techniques to improve detection outcomes in downstream processes [22].…”

Microfluidic and mathematical modeling of aquatic microbial communities

“…Microbial growth depends sensitively on temperature. Temperature control has been developed previously in microfluidic platforms for biochemical reactions that require thermal cycling [50][51][52][53], such as the polymerase chain reaction (PCR), for the preparation of thermal-sensitive materials [54], and for the investigation of temperature-dependent biological processes such as oocyte membrane permeability in oocyte cryopreservation [55,56]. Heating and cooling, as well as sensing and feedback control, are important components of temperature control [57][58][59].…”

Passively driven microfluidic device with simple operation in the development of nanolitre droplet assay in nucleic acid detection

“…Additionally, other functional devices, such as simple pumping systems, valves and automatic systems, can easily be combined with the microfluidic system, allowing further related analytic developments 18 . Due to several advantages, including reduced consumption of clinical sample reagents, higher reaction efficiency due to low thermal mass inertia with rapid heat transfer, portability, automation ability and reduced human operating error 19 – 21 , the MF system is especially of interest for nucleic acid amplification technology in the POCT field 22 – 24 . Many studies have incorporated nucleic acid detection devices into microfluidic systems.…”