Abstract: OBJECTIVE To establish a rapid detection method for human respiratory syncytial virus A (HRSV-A) based on reverse transcription loop-mediated isothermal amplification (RT-LAMP) combined with fluorescent probe technology, thereby providing technical support for on-site rapid screening. METHODSSpecific primers and probes were designed targeting the conserved region of the M gene of HRSV-A. A 25 μl reaction system was established and optimized, and then incubated at a constant temperature of 64 ℃ for 30 minutes, while the nucleic acid amplification process was monitored in real time by fluorescence. The analytical sensitivity was evaluated by absolute quantification of recombinant plasmids through droplet digital polymerase chain reaction (PCR), and the specificity was verified through standards of 11 common respiratory pathogens. Furthermore, a commercial HRSV typing kit (RT-PCR method) and the RT-LAMP fluorescent probe method established in this study were adopted for parallel detection of 160 clinical samples to comprehensively evaluate their diagnostic performance. RESULTSThe detection limit of the RT-LAMP fluorescent probe method established in this study for HRSV-A was 500 copies/ml. The specificity results showed that it only produced specific amplification for HRSV-A, with no cross-reactivity with HRSV-B or other respiratory pathogens. Clinical sample validation demonstrated a total concordance rate of 100.00%, with both sensitivity and specificity at 100.00%, and a Kappa value of 1.000 (95% CI: 1.000-1.000) compared to the commercial HRSV typing kit. The total detection time was approximately 22.53 minutes. CONCLUSIONS This study successfully developed a rapid detection method for HRSV-A based on RT-LAMP fluorescent probe technology. This method is rapid, sensitive, and specific, capable of completing detection within 30 minutes, and is suitable for various scenarios of on-site rapid screening, including fever clinics and pediatric outpatient departments.