Pepsin　serves　as　a　potential　biomarker　for　laryngopharyngeal　reflux　disease　(LPRD).　The　reported　methods　for　detecting　pepsin　still　have　limitations,　such　as　long　reaction　times　and　low　sensitivity.　Therefore,　it　is　crucial　to　develop　a　rapid,　simple　and　sensitive　detection　method.　In　this　study,　a　fluorescence　immunochromatographic　test　strip　was　designed　for　point-of-care　diagnostics　of　LPRD.　To　obtain　optimal　fluorescence　immunochromatographic　conditions,　key　experimental　factors,　including　microsphere　particle　size,　binding　pH,　and　resuspension　dilution　ratio,　were　systematically　evaluated.　The　optimal　fluorescent　microsphere　size　was　determined　to　be　300　nm,　the　most　suitable　reaction　pH　was　8.0,　and　the　ideal　resuspension　solution　to　storage　solution　ratio　was　1　:　4.　Under　these　optimal　conditions,　the　sensor　had　a　linear　range　of　2.5-100.0　ng　mL-1　and　a　detection　limit　of　1.9　ng　mL-1.　The　relative　standard　deviation　was　below　15%　for　both　intra-　and　inter-batch　reproducibility.　Under　accelerated　damage　conditions　at　37　degrees　C,　the　test　value　of　the　test　strip　could　be　kept　basically　unchanged　for　14　days.　There　was　no　cross-reactivity　of　the　test　strip　with　pepsinogen　I,　pepsinogen　II　and　other　interferents.　In　the　actual　sample　detection,　the　recovery　rate　was　between　85%　and　115%.　Furthermore,　the　proposed　method　was　demonstrated　to　be　green　and　to　exhibit　strong　applicability　based　on　the　evaluation　using　the　AGREEprep　and　the　BAGI.　In　addition,　the　method　has　the　advantages　of　high　sensitivity,　accuracy,　greenness　and　applicability,　which　can　meet　the　needs　of　rapid　quantitative　on-site　detection.