Rapid 14-Minute Detection: Development and Application of a Portable Nucleic Acid Rapid Testing Device for Primary Care

Overview

Rapid identification of respiratory pathogens is essential for selecting the appropriate treatment, especially in primary care clinics and developing areas with limited medical resources. I developed a portable nucleic acid testing device that combines recombinase polymerase amplification (RPA), CRISPR-Cas12a detection, and smartphone-based signal analysis. The test includes a 6-minute RPA amplification step followed by a 6-minute CRISPR detection step, giving a total turnaround time of 14 minutes, which is much faster than the conventional qPCR method. The device can run five reactions simultaneously to detect influenza A, influenza B, Mycoplasma pneumoniae, an internal control, and a no-template control. Fluorescence signals are captured by a smartphone and analyzed by a custom app that automatically reports positive or negative results, reducing user bias. The system runs on a portable power bank and does not require laboratory equipment. It achieved a limit of detection of 1 copy/μL and showed results comparable to qPCR when tested with clinical samples. Overall, this platform provides a fast, low cost solution for point-of-care nucleic acid testing and has potential applications in primary healthcare settings.