Hepatitis B virus (HBV) infection is a public health problem in the world. Even though the association between maternal HBV infection and adverse pregnancy outcomes has been extensively discussed, limited and conflicting results still led us confusing. To clarify the association between maternal HBV infection and adverse pregnancy outcomes, we conducted a retrospective cohort study which include 65,257 pregnant women who performed non-invasive prenatal screening (NIPS) in second-trimester from July 2015 to Nov 2019 in Suzhou, China. The participants were divided into two groups according to their status of HBV infection: 63,591 pregnant women with HBV un-infected (as control) and 1,666 pregnant women infected with HBV (exposure group). Meanwhile, eight types of adverse pregnancy outcomes in history and twelve prenatal screening results in the second-trimester of current pregnancy were investigated by estimating and adjusting their risk ratios (aRR) for women between HBV infected and uninfected using multivariate logic regression. Our results suggested that women infected with HBV have higher risks on the biochemical pregnancy (aRR: 1.54, 95%CI:0.97-2.32, p=0.048), extrauterine pregnancy (aRR: 1.36, 95%CI: 1-1.82, p =0.043) and three kinds of screen result for fetal Down's syndrome (DS) than women with HBV un-infected. The risks of increased thickness of nuchal translucency or nuchal fold (I-NTF) is 123% higher (aRR 2.23, 95%CI 1.17-3.82, p = 0.007) and the risk of DS positive possibility in multiples serum markers (MSM) and NIPS respectively increased 22% (aRR: 1.22, 95%CI:1.07-1.39, p=0.003) and 88% (aRR: 1.88, 95%CI: 1.07-3.06, p= 0.018). Finally, the true positive risk of DS in NIPS is 100% higher (aRR: 2.0, 95%PI: 1.02-3.18, p= 0.017) in women with HBV infected than uninfected. Importantly, the fetal risk of DS in women with HBV infected conformed by four different detection methods and their analysis results were supported by all adjusted models. Our results provide robust evidence to support that maternal HBV infection was an independent risk factor to associate with fetal DS, biochemical pregnancy and extrauterine pregnancy. Although the causality has not yet to be determined, our study provides an opportunity to gain deep insights into the impact of maternal HBV infection on pregnancy and the potential pathogenic mechanism of DS and abnormal pregnancy.