Challenges in the Development and Implementation of Point-of-Care Handheld qPCR Devices

Polymerase chain reaction (PCR) is a highly sensitive and accurate technique for amplifying and detecting nucleic acids (NA). Its extensive applications in clinical diagnostics, environmental monitoring, agricultural, and forensic science highlight the critical need for translating this laboratory-based technique into a point-of-care (POC) quantitative PCR (qPCR) platform. The transition to POC qPCR faces significant challenges, including the need to streamline sample preparation, miniaturize devices without compromising their analytical performance, and achieve cost-effective and standardized platforms suitable for use by non-experts. This review discusses the challenges and approaches involved in developing simplified sample-preparation methods and miniaturized qPCR devices suitable for on-site implementation. The speed, efficiency, and simplicity of recent sample preparation methods that utilize microfluidics and nanoparticles are highlighted as pivotal innovations for NA extraction. Approaches for developing thermal cyclers and NA detection methods, emphasizing the hurdles and limitations that impede effective device miniaturization and field implementation, are then examined. This review also discuss strategies to promote widespread adoption of POC qPCR devices, with a focus on cost reduction and standardization as critical enabling factors. Finally, the key opportunities to address these challenges are outlined through practical considerations, thereby supporting the development of rapid, precise, handheld, and low-cost POC qPCR devices.