Construction of dPCR and qPCR integrated system based on commercially available low-cost hardware

Abstract

Fluorescent quantitative PCR (qPCR) and digital PCR (dPCR) are two mainstream nucleic acid quantification technologies. However, commercial dPCR and qPCR in-struments have a low integration, a high price, and a...

A sample-to-answer DNA detection microfluidic system integrating sample pretreatment and smartphone-readable gradient plasmonic photothermal continuous-flow PCR

“…As for ultra-sensitive quantification of DNA at lower concentrations (dozens of copies or even a single copy), studies have shown that ordinary fluorescence PCR is inherently unreliable, so the digital PCR method was specially invented to meet the needs of absolute quantification, making up for the inadequacy of real-time PCR. 22,52 Comparative analysis and outlook…”

“…28 Therefore, an affordable smartphone imaging device can obtain the fluorescence intensities of all cycles for sensitive and timely DNA analysis, which is superior to qualitative electrophoresis and dedicated multi-fiber detection or multi-zone fluorescence imaging. 22,29,30 As a proof of concept, we built a 3-D printed prototype and evaluated the performance including nucleic acid extraction efficiency and quantification ability with hepatitis B virus DNA (HBV-DNA) spiked plasma samples. The results show that this study can complete 69% DNA extraction from a 100-μL plasma sample within 14 minutes, and achieve real-time fluorescence amplification of a 40-μL PCR reagent within 23 minutes, with a quantitative range of up to 6 orders of magnitude.…”

“…21 Zhenqing Li et al integrated the continuous-flow PCR (CF-PCR) in 151 seconds and electrophoresis in 223 seconds into a polycarbonate (PC) microdevice. 9 Wenming Wu's team successively proposed a 3-D helical tubular CF-PCR device 18 and a scheme of moving a PCR or digital PCR chip between multiple heating zones, 22 avoiding dynamic thermal control. These studies have accelerated some (but not all) aspects of PCR detection, and there is room for improvement in sample preparation and content characterization.…”

“…28 Therefore, an affordable smartphone imaging device can obtain the fluorescence intensities of all cycles for sensitive and timely DNA analysis, which is superior to qualitative electrophoresis and dedicated multi-fiber detection or multi-zone fluorescence imaging. 22,29,30…”

Development of a low-cost multi-channel nucleic acid detection PCR instrument and clinical detection application of COVID-19

“…The detection of novel coronavirus is an important part of controlling the epidemic. Therefore, it is imperative to develop a test instrument that is easy to operate and quick to measure [ 21 , 22 ].…”