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This study determined the effect of hepatitis B virus (HBV) replication in peripheral blood mononuclear cell (PBMC) from HBsAg-positive mothers on HBV intrauterine transmission.A total of 150 HBsAg-positive mothers and their neonates were recruited in this study.Within 24 h after birth,HBV serological markers,serum HBV DNA,PBMC HBV relaxed circular DNA (rcDNA),and covalently closed circular DNA (cccDNA) were measured in the HBsAg-positive mothers and their neonates before passive-active immune prophylaxis.The relationship between HBV replication in PBMC and HBV intrauterine transmission was examined through Chisquare test and logistic regression.The rate of HBV intrauterine transmission was 8.00％ (12/150) in the 150 neonates bo to HBsAg-positive mothers.The positivities of PBMC HBV rcDNA and cccDNA in the HBsAg-positive mothers were 36.67％ (55/150) and 10％ (15/150),respectively.Mateal PBMC HBV cccDNA was a risk factor of HBV intrauterine transmission (OR =6.003,95％ CI:1.249-28.855).Mateal serum HBeAg was a risk factor of PBMC HBV rcDNA (OR =3.896,95％ CI:1.929-7.876) and PBMC HBV cccDNA (OR =3.74,95％ CI:1.186-11.793) in the HBsAg-positive mothers.Administration of hepatitis B immune globulin was a protective factor of PBMC HBV cccDNA (OR =0.312,95％ CI:0.102-0.954) during pregnancy.The positivity of PBMC HBV rcDNA was related to that of cccDNA in the HBsAg-positive mothers (x2=5.087,P =0.024).This study suggests that PBMC is a reservoir of HBV and an extrahepatic site for virus replication and plays a critical role in HBV intrauterine transmission.