Development of digital polymerase chain reaction

abstract

English

During the last 20 years, microfluidics has proven to be a powerful nucleic acid (NA) analysis tool in applications such as point-of-care diagnostic methods, which critically depended on miniaturization. From simple systems they evolved into complex devices integrating sample processing, NA amplification and product detection using samples with an ultra-small volume, resulting in inexpensive screening. There are even techniques which cannot be conducted without microfluidics, such as digital polymerase chain reaction (dPCR) developed in 1999. We demonstrate a concept of droplet real-time PCR (qPCR) which will be developed into dPCR. For droplet real-time detection, the amplification is conducted in 0.3-µL master mixture droplet containing target gene encapsulated in 2 µL of mineral oil. For digital detection, a chip-based system with microwell sample dispersion will be developed. The 3 µL of master mix solution with DNA will be pipetted on a silicone chip and covered with a hydrophobically treated glass. We improved the features of both devices where the experimental setup for qPCR and dPCR is the same, except the detectors. Our detection system is more economic thanks to the use of a recyclable microchip, lower consumption of components and shortened detection time, which makes it attractive for point-of-care applications.

Digital PCR, miniaturization, microchip technology

Gaňová Martina, Neužil Pavel

26. 2. 2021

TAYLOR & FRANCIS LTD

European Biotechnology Congress 2020 Abstracts, ABINGDON

1314-3530

Biotechnology and Biotechnological Equipment

35

sup1

United Kingdom of Great Britain and Northern Ireland

S62

S128

67

https://doi.org/10.1080/13102818.2020.1871545