Objective　To　develop　a　rapid,　highly　sensitive　quantitative　method　for　detecting　P24　antigen　based　on　near-infrared　fluorescent　microsphere　immunochromatography.　Methods　First,　we　prepared　a　lateral　flow　assay　test　strip,　and　labeled　the　detection　antibody　using　a　fluorescent　microsphere.　Second,　we　optimized　the　antibody　labeling　conditions.　Third,　we　optimized　the　detection　conditions.　Fourth,　we　created　a　working　curve.　Fifth,　we　conducted　a　methodological　assessment　of　the　established　fluorescent　microsphere　immunochromatography　method.　Sixty-six　clinical　samples　were　tested,　and　we　compared　the　established　fluorescent　microsphere　immunochromatography　with　the　quantitative　ELISA　method.　Results　According　to　the　working　curve,　the　detection　limit　of　the　method　is　3.4　pg/mL,　and　the　detection　range　is　3.4　pg/mL　to　10　ng/mL.　The　average　intra-assay　recovery　was　99.6%,　and　the　Coefficient　of　Variation　(CV)　was　5.4%-8.6%;　the　average　inter-assay　recovery　was　97.3%,　and　the　CV　was　8.5%-11%.　The　detection　rate　of　fluorescent　microsphere　immunochromatography　was　higher　than　ELISA　method,　and　had　a　good　correlation　with　ELISA.　Conclusion　The　P24　antigen　quantitative　detection　method　based　on　near-infrared　fluorescent　microsphere　immunochromatography　has　the　advantages　of　rapid　detection,　high　sensitivity,　and　wide　detection　range;　thus,　it　is　suitable　for　early　clinical　diagnosis　and　continuous　monitoring　of　AIDS.