Low-Cost Battery-Powered and User-Friendly Real-Time Quantitative PCR System for the Detection of Multigene

Abstract

Real-time polymerase chain reaction (PCR) is the standard for nucleic acid detection and plays an important role in many fields. A new chip design is proposed in this study to avoid the use of expensive instruments for hydrophobic treatment of the surface, and a new injection method solves the issue of bubbles formed during the temperature cycle. We built a battery-powered real-time PCR device to follow polymerase chain reaction using fluorescence detection and developed an independently designed electromechanical control system and a fluorescence analysis software to control the temperature cycle, the photoelectric detection coupling, and the automatic analysis of the experimental data. The microchips and the temperature cycling system cost USD 100. All the elements of the device are available through open access, and there are no technical barriers. The simple structure and manipulation allows beginners to build instruments and perform PCR tests after only a short tutorial. The device is used for analysis of the amplification curve and the melting curve of multiple target genes to demonstrate that our instrument has the same accuracy and stability as a commercial instrument.

Fraud in Animal Origin Food Products: Advances in Emerging Spectroscopic Detection Methods over the Past Five Years

“…In recent applications, entirely satisfactory limit of detection values were achieved [276,277]. Efforts have been made to develop low-cost and user-friendly PCR devices with accuracy and stability comparable to commercial alternatives [278]. Commercial PCR-based assays designed for the detection and quantitative authentication of animal species in a specific dairy product are also available in the market [279,280].…”

Cost-Effective Multiplex Fluorescence Detection System for PCR Chip

“…Although PCR is a simple and straightforward diagnosis method, the operation is mainly carried out in certain facilities such as in hospitals or central laboratories due to portability and cost limitations, which can be disadvantageous in emergencies [1,[21][22][23][24][25][26]. For example, there are cases where administration of Tamiflu within 48 h of initial symptom observation is critical for influenza patients [27].…”

“…However, it is difficult to meet the 'ASSURED' criteria and use the equipment at resource-limited settings since it incorporates highly a sensitive fluorescence detection method, increasing the cost to 10-fold that of a conventional PCR platform [33][34][35][36][37]. To overcome these limitations, various microfluidic chips and low-cost fluorescence detection methods have been reported [23,26,[38][39][40][41]. El-Tholoth et al (2021) used real-time reverse transcriptase loop-mediated isothermal amplification (qRT-LAMP) to lower the thermal cycle cost, but the cost of fluorescence detection remained an issue because they used a universal serial bus (USB) fluorescence microscope.…”

Cost-Effective Multiplex Real-Time PCR Chip System Using Open Platform Camera

“…As they are usually expensive, only large hospitals or laboratories can afford them. However, nowadays it is very important to quickly and accurately diagnose viruses for infectious diseases, requiring a PCR device capable of point-of-care test (POCT) [1,2]. For POCT devices, miniaturization and cost reduction are essential, which is what most PCR chips aim for [1][2][3][4].…”