OBJECTIVE To test the hypothesis that concentration of amniotic fluid alpha-fetal protein (AFAFP) is increased in thalassemia fetus. METHODS A total of 135 cases of amniocentesis admitted from July 2013 to December 2014 were included in this study. Among them 98 cases of normal fetuses were assigned into control group and 37 cases of thalassemia fetus were included as thalassemia fetus group. Alpha-fetoprotein levels detected by enzyme linked immunosorbent assay and the alpha-fetoprotein concentration were compared between the two groups. There is no significant difference in gestational age between the two groups. RESULTS 1. AFP concentration in thalassemia fetus group was significantly higher than that of normal control group [(1541.65 ± 734.78) μg/mL vs. (2728.84 ± 1539.97) μg/mL], and amniotic fluid AFP concentration was related to fetal thalassemia. 2. AFAFP concentration in pure α-thalassemia fetus was higher than that of β-thalassemia fetus or mixed α- and β-thalassemia fetus, but the difference was not significant. CONCLUSIONS Concentration of amniotic fluid alpha-fetal protein is increased in thalassemia fetus. AFP concentration in α-thalassemia fetus was higher than that of β-thalassemia or mixed α- and β-thalassemia fetus but difference was not significance. Further studies are needed to explore the possible correlation between Down syndrome and biochemical markers of thalassemia.