The association between the embryo quality, clinical characteristics, miRNAs (secreted by blastocysts in the culture medium) and pregnancy outcomes has been well-established. Studies on prediction models for pregnancy outcome, using clinical characteristics and miRNA expression, are limited. We aimed to establish the prediction model for prediction of pregnancy outcome of woman after a fresh cycle of Day 5 single blastocyst transfer (Day 5 SBT) based on clinical data and miRNA expression. A total of 86 women, 50 with successful pregnancy and 36 with pregnancy failure after fresh cycle of Day 5 SBT, were enrolled in this study. All samples were divided into training set and test set (3:1). Based on clinical index statistics of enrolled population and miRNA expression, the prediction model was constructed, followed by validation of the prediction model. Four clinical indicators, female age, sperm DNA fragmentation index, anti-mullerian hormone, estradiol, can be used as independent predictors of pregnancy failure after fresh cycle of Day 5 SBT. Three miRNAs (hsa-miR-199a-3p, hsa-miR-199a-5p and hsa-miR-99a-5p) had a potential diagnostic value for pregnancy failure after Day 5 SBT. The predictive effect of model combining 4 clinical indicators and 3 miRNAs (area under the receiver operating characteristic curve, AUC = 0.853) was better than models combining single 4 clinical indicators (AUC = 0.755) or 3 miRNAs (AUC = 0.713). Based on 4 clinical indicators and 3 miRNAs, a novel model to predict pregnancy outcome in woman after fresh cycle of Day 5 SBT has been developed and validated. The predictive model may be valuable for clinicians to make the optimal clinical decision and patient selection.