An automated portable LAMP-based centrifugal microfluidic system for nucleic acid detection of multiple pathogens in feline upper respiratory disease.

Abstract

A portable centrifugal microfluidic nucleic acid analyzer tailored for point-of-care diagnostics was developed. The system consists of a main unit and a sector-shaped microfluidic chip driven by centrifugal force. By incorporating branched microchannels into the chip design, the system enables simultaneous detection of multiple targets from a single sample. The sample flows sequentially through the sample reservoir, dissolution chamber, and metering structure before reaching the reaction chamber. Under the presence of specific primers, the analyzer performs loop-mediated isothermal amplification (LAMP) by heating the chip. Fluorescence signals are captured by the optical detection module and displayed in real-time. We designed highly specific primers targeting six common feline upper respiratory disease (FURD) pathogens and extensively validated the device's specificity and sensitivity. The system demonstrated a low detection limit (10-100 copies/μL) and successfully identified 72 clinically positive samples, achieving 100% agreement with conventional PCR. Weighing only 5 kg, the analyzer can accurately detect and identify six FURD-related pathogens from a single sample within 30 min. This method enables a shift from single-target to multitarget detection and promotes the broader adoption of decentralized testing. It offers an efficient and convenient solution for rapid and accurate pathogen detection, particularly in pet hospitals and veterinary clinical diagnostics.