Design and fabrication of microfluidic chip and integrated nucleic acid rapid diagnosis device

• Experimental results demonstrate high sensitivity (94.1 %, 32/34), specificity (97.7 %, 42/43), and accuracy (96.1 %, 74/77), with a detection limit of 10² copies/μL. • The device features low production costs (approximately $69.1), low per-test expenses, and a portable design (2 kg), facilitating field deployment. • A low-cost microfluidic chip and diagnostic equipment is designed and fabricated using a mini CNC milling machine and a 3D printer. • The diagnosis can complete without the assistance of external equipment and manual operation. African Swine Fever (ASF) poses a severe threat to the global pig husbandry. In the absence of an effective vaccine, developing rapid and accurate diagnostic methods is crucial for epidemic control. This study designed a low-cost microfluidic chip and integrated nucleic acid rapid detection device (INARDD) based on colorimetric principles combined with magnetic bead based nucleic acid extraction technology. Manufactured using micro computer numerical control machine tools (CNC) and 3D printing, the device fully automates the entire process include nucleic acid extraction, transfer, loop-mediated isothermal amplification (LAMP), and detection without requiring external equipment or manual intervention. Experimental results demonstrate high sensitivity (94.1 %, 32/34), specificity (97.7 %, 42/43), and accuracy (96.1 %, 74/77), with a detection limit of 10² copies/μL. The device features low production costs (approximately $69.1), low per-test expenses, and a portable design (2 kg), facilitating field deployment. This study provides an economical and efficient solution for rapid on-site diagnosis of African swine fever, holding significant implications for enhancing epidemic control and ensuring meat safety. It also offers technical reference for developing portable diagnostic systems for other infectious diseases. Schematic diagram of the microfluidic chip diagnostic process.