Abstract: Objective　To develop rapid amplified luminescent proximity homogeneous immunoassay (AlphaLISA) reagent for the detection of human follicle stimulating hormone(hFSH).  Methods　Two monoclonal anti-hFSH antibodies were used to develop the AlphaLISA reagent for the detection of hFSH. One was biotinylated，and the other was coated on AlphaLISA acceptor beads. The reagent also contained donor beads coated with streptavidin. The optimal test conditions and analytical performance of the method were evaluated.  Results　The analytical sensitivity and functional sensitivity of AlphaLISA for the detection of hFSH were 0.09 and 0.12 U/L. The linear measurement range of the developed reagent was 0.09-192.00 U/L. The intra- and inter-assay coefficients of variation (CV) were 4.2%-7.1% and 7.5%-8.3%，which were all<10.0%，respectively. The ratios of cross-reactivity were 0.16%，0.08% and 0.02% with human luteinizing hormone(hLH)，human thyroid stimulating hormone(hTSH) and human chorionic gonadotropin(hCG)，respectively. The results of 100 samples′ detection by this reagent showed good correlation coefficient with those of SIEMENS Immulite 2000 follicle stimulating hormone kit by chemiluminescent immunoassay (CLIA，r=0.979，P<0.001).  Conclusions　The developed hFSH AlphaLISA reagent in this study meets the requirement of clinical determination，and is a valuable test for clinical application.