Background: The detection of hepatitis B surface antigen (HBsAg) positivity in pregnant women before delivery is crucial to preventing mother-to-child transmission of hepatitis B virus. Aim: This study aimed to evaluate the status and rate of testing for HBsAg, the rate of HBsAg positivity and the distribution of this rate by age and to evaluate changes in the HBsAg positivity rate in pregnant women over the study period. Methods: We conducted a multicentre, cross-sectional, descriptive study covering the period January 2005 to June 2019. The 15 years of data for 2 145 668 pregnant women from 27 provinces in all 7 regions of Turkey were collected and recorded in the Excel database before the statistical analysis. Results: We found 1 012 593 (47.1%) pregnant women had been tested for HBsAg and 11 471 (1.1%) were HBsAg-positive. Overall, 97% of the HBsAg positive women were born before 1998, the year that national HBV vaccination was launched in Turkey. The rate of HBsAg positivity in that group was 1.1% compared with 0.3% among women born after 1998. Conclusion: The downward trend in the HBsAg positivity rate among pregnant women in younger age groups, especially in those born after universal HBV vaccination was inaugurated, and the low rate of testing for HBsAg during pregnancy pronounced in our study.