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Objective: To study the value ofα-fetoprotein (α-FP),β-human chorionic gonadotropin (β-HCG), inhibin A, and uncojugated estriol (UE3) at the second trimester for early screening of preeclampsia. Methods: This cohort study was performed on 1713 consecutive pregnant women with gestational age from 15 to 18 weeks. We measured the serum levels ofα-FP, β-HCG, inhibin A, and UE3 and they were followed-up for 48 h after delivery. Results: A total of 1713 women were recruited and through the study 151 women (8.8％) were diagnosed with preeclampsia (n=123, 21.5％ in the pregnant women with high risk, n=28, 2.5％ in the pregnant women with low risk; P<0.0001). The area under receiver operating characteristic curve was estimated to be 2.39 MOM forα-FP, 2.84 MOM forβ-HCG, 1.92 MOM for inhibin A, and 0.77 MOM for UE3. The best cutoff value was specifically considered for each marker that was determined a positive predictive value ranged 19.60％ forα-FP, 17.62％ for β-HCG, 18.33％ for inhibin A and 14.88％ for UE3, a negative predictive value ranged 95.63％ for α-FP, 93.89％ for β-HCG, 94.28％ for inhibin A and 93.57％ for UE3, and an area under receiver operating characteristic curve ranged 0.70 forα-FP, 0.63 forβ-HCG, 0.65 for inhibin A and 0.61 for UE3. Conclusions: This study reveals that the new cutoff values are more valuable in screening preeclampsia. Although these markers have a low positive predictive value, they present a high negative predictive value. Also, the combination ofα-FP,β-HCG and inhibin A and the combination ofα-FP and inhibin A have the highest value in screening preeclampsia.