Objective · To understand the cognition, attitude, willingness and demand for non-invasive prenatal testing (NIPT) of pregnant women. Methods · A total of 852 pregnant women were enrolled, including 318 pregnant women with low risk of Down syndrome screened in Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, and 534 pregnant women with high risk of Down syndrome screened in several hospitals in Shanghai who further went to Xinhua Hospital, Shanghai Jiao Tong University School of Medicine for NIPT to confirm the diagnosis of Down syndrome screening. The information about the cognition, attitude, willingness and demand for NIPT was collecteda standard questionnaire. Results · A total of 760 questionnaires were collected, of which 728 ones were valid, with an effective questionnaire rate of 85.45%. The proportion of pregnant women with ideal cognition level was 51.24%. However, 83.10% of pregnant women held a positive attitude towards the promotion of NIPT. There were 79.54% of pregnant women considering it necessary to provide genetic counseling before NIPT. Conclusion · It is of great significance to strengthen the prenatal propaganda and education and pre-test genetic counseling so as to improve the cognitive level of pregnant women and make rational of NIPT.