A　high-throughput,　simple　and　rapid　chemiluminescence　enzyme　immunoassay　(CLEIA)　was　developed　for　the　clinical　determination　of　progesterone　in　human　serum,　and　the　luminol-hydrogen　peroxide　was　used　as　chemiluminescence　system　catalyzed　by　horseradish　peroxidase　(HRP).　The　solid　phase　of　anti-progesterone　antibody　was　prepared　through　the　immunoreaction　between　anti-progesterone　polyclonal　antibody　with　donkey　anti-rabbit　IgG,　i.　e.,　second　antibody,　which　had　been　physically　absorbed　on　the　wells　of　polystyrene　microplate　and　was　used　as　a　universal　solid　phase.　The　effect　of　several　factors,　such　as　the　dilution　ratios　of　the　immunoreagents,　the　chemiluminescent　substrate,　the　chemiluminescence　reaction　time　and　incubation　condition　were　examined　and　optimized.　The　optimal　dilutions　of　anti-progesterone　antibody　and　HRP-progesterone　conjugate　were　1:　10000　and　1:　15000,　respectively.　The　substrate　U　was　chosen　and　the　luminescence　was　determined　after　10　min　incubation.　The　immunoreaction　was　incubated　in　water　bath　at　37　degrees　C　for　1　h.　The　assay　was　evaluated　with　sensitivity　as　low　as　0.08　mu　g/L.　The　RSD　was　less　than　15%　for　both　intra-　and　inter-assay　precision.　The　recoveries　of　three　different　spiked　concentration　samples　were　101%,　101%　and　94.4%,　respectively.　This　proposed　method　has　been　successfully　applied　to　the　clinical　evaluation　of　progesterone　in　36　human　sera.　The　results　showed　a　good　correlation　with　the　commercially　available　radioimmunoassay　kit　with　a　correlative　coefficient　of　0.9502.