Isothermal Recombinase Polymerase Amplification (RPA) of E. coli gDNA in Commercially Fabricated PCB-Based Microfluidic Platforms.

Maria Georgoutsou-Spyridonos,Angeliki Tserepi,Dimitrios C. Mastellos,Myrto Filippidou,Stavros Chatzandroulis,Georgia D. Kaprou

doi:10.3390/mi12111387

Maria Georgoutsou-Spyridonos, Angeliki Tserepi + Show 4 more

Open Access

https://doi.org/10.3390/mi12111387

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Abstract

Printed circuit board (PCB) technology has been recently proposed as a convenient platform for seamlessly integrating electronics and microfluidics in the same substrate, thus facilitating the introduction of integrated and low-cost microfluidic devices to the market, thanks to the inherent upscaling potential of the PCB industry. Herein, a microfluidic chip, encompassing on PCB both a meandering microchannel and microheaters to accommodate recombinase polymerase amplification (RPA), is designed and commercially fabricated for the first time on PCB. The developed microchip is validated for RPA-based amplification of two E. coli target genes compared to a conventional thermocycler. The RPA performance of the PCB microchip was found to be well-comparable to that of a thermocycler yet with a remarkably lower power consumption (0.6 W). This microchip is intended for seamless integration with biosensors in the same PCB substrate for the development of a point-of-care (POC) molecular diagnostics platform.

Highlights

The accommodation of conventional laboratory processes in microfluidic platforms fabricated using well-established microfabrication technology has drawn great attention and led to the development of the so-called lab-on-a-chip (LOC) devices
Escherichia coli are a common, large, and diverse group of bacteria found in the environment, food, and intestines of people and animals
The recombinase polymerase amplification (RPA) was selected as the amplification method because it is an isothermal one, avoiding thermocycling, while in addition, it can reduce analysis time much below 60 min

Summary

Introduction

The accommodation of conventional laboratory processes in microfluidic platforms fabricated using well-established microfabrication technology has drawn great attention and led to the development of the so-called lab-on-a-chip (LOC) devices. Typical advantages of microfluidic systems include the possibility to use very small quantities of expensive reagents and scarce samples, to perform high-resolution, precise, and sensitive detection, and to reduce the analysis time and cost [1]. Such systems are capable of performing a great variety of laboratory processes, such as sample purification and enrichment [2,3,4], reagent mixing [5,6], ultra-fast thermal cycling required in many biochemical reactions [7], as well as the detection of reaction products [8,9], which is of utmost importance. LOC devices typically integrate microfluidic components, electrical driving circuits, and sensors into the same, usually hybrid, platform [8,10,11,12].

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