Objective:To explore advantages of high-throughput sequencing of cell-free DNA from maternal peripheral blood for detection of chromosomal aneuploidies in prenatal diagnosis. Methods:A total of 1 000 maternal peripheral blood samples from gravidas with more than 13 weeks of pregnancy were collected,and the cell-free DNA in the peripheral blood samples were detected using high-throughput sequencing to determine chromosomal aneuploidy. The cases of chromosomal aneuploidies and euploidies were validated by traditional chromosomal karyotype analysis and clinical follow-up of babies after the birth. Results:In the analyses of 1 000 cases,one case failed. The results of the remaining 999 cases indicated 18 cases of abnormalities,including 12 cases of trisomy 21,two cases of trisomy 18,one case of trisomy 13,one case of X monomers,one case of trisomy X,one case of XXY. From 12 cases of trisomy 21,there was one case of gemellary pregnancy,and the results of fetal karyotype analysis showed that one fetal was euploid and the other fetal was trisomy 21. One case of trisomy X was validated as euploid by karyotype analysis and the remaining detection results were consistent to those with karyotype analysis. The total positive detection rate of trisomy 21,trisomy 18,trisomy 13 and sex chromosome aneuploidy was 94.44%. No abnormal signs of chromosome aneuploidies (trisomy 21,trisomy 18 and trisomy 13) were found in follow-up of face and physical development of babies after the birth for all cases with negative results,and there were no false-negative cases. Conclusion:High-throughput sequencing of cell-free DNA from maternal peripheral blood has high accuracy for detecting fetal trisomy 21,trisomy 18 and trisomy 13. For sex chromosome aneuploidy detection,measurement indicators in this study should be further improved.