新生儿外周血单个核细胞乙型肝炎病毒DNA检测的临床意义

Clinical significance of detecting neonatal peripheral blood mononuclear cells infected by HBV

目的探讨HBsAg阳性产妇新生儿外周血单个核细胞(PBMC)乙型肝炎病毒(HBV)感染率、影响因素及检测的临床意义。方法50例HBsAg阳性产妇和其新生儿为研究组,另18例血清乙肝标志物均阴性者为对照组。采用套式聚合酶链反应(n-PCR)法分别检测产妇、新生儿血清及PBMCHBV-DNA。结果HBsAg阳性母亲血清及PBMCHBV-DNA检出率分别为60·0%和40·0%;其新生儿血清及PBMCHBV-DNA阳性率分别为46·0%及30·0%;其中仅血清阳性10例,仅PBMC阳性2例,血清和PBMC均阳性13例。对照组母儿血清及PBMC中未检出HBV-DNA。新生儿PBMCHBV-DNA阳性率在母血清HBeAg、HBV-DNA阳性组高于阴性组(P<0·05,P<0·01);母PBMCHBV-DNA阳性组高于阴性组(P<0·01);母血清和PBMCHBV-DNA均阳性组高于仅血清阳性组(P<0·01);新生儿血清HBV-DNA阳性组高于阴性组(P<0·05)。结论HBsAg阳性母亲新生儿HBV-DNA感染率为30·0%;其感染率与母、儿血清病毒血症水平及孕妇PBMCHBV-DNA状态有关,检测出新生儿PBMCHBV-DNA具有重要临床意义。

Objective To understand the HBV infection rate of peripheral blood mononuclear cells (PBMCs) from fetuses of HBsAg positive mothers, associated risk factors and to explore the clinical significance of detecting HBV infected PBMCs.Methods Sixty eight pregnant women who were delivered at the First Hospital of Xi'an Jiaotong University, China from August 1995 to February 1997, and their newborns were studied. They were divided into two groups according to their status of HBV serological markers. The study group included 50 cases who were serum HBsAg positive and 18 cases without any HBV serum markers served as control group. All these cases had no symptoms of hepatitis, high risk premature labor, premature delivery and hypertensive disorder complicating pregnancy. Age and gestational age were matched in two groups. Blood samples (5 mL) were taken from the peripheral vein of pregnant women before delivery and from newborns within 24 h after birth, before inoculation of HBV vaccine (HBVac) and injection of hepatitis B immunoglobulin (HBIG). PBMCs were isolated. The sera and PBMCs were stored at -80℃. HBV- DNA in serum and PBMCs were detected with nested polymerase chain reaction (n-PCR). Two pairs of oligonucleotide primers, the outer primer pair for first PCR and inner primer pair for second PCR, designed according to region S of HBV genome were synthesized by Shanghai Cell Biology Institute of Chinese Academy of Science. Results The detection rate of HBV-DNA in serum and PBMCs from HBsAg positive pregnant women was 60.0%(30/50)and 40.0%(20/50), respectively. The detection rate of HBV-DNA in serum and PBMCs from newborns of HBsAg positive pregnant women was 46.0%(23/50) and 30.0% (15/50), respectively. Ten newborns were HBV- DNA positive in serum only, 2 were positive in PBMCs only and 13 were positive in both serum and PBMCs. In the control group, HBV- DNA was not detected in PBMC nor in serum. The positive rate of HBV-DNA in PBMCs of newborns was significantly higher in the group of mothers who were HBV-DNA or HBeAg positive in serum( P <0.05, P <0.01);the positive rate was significantly higher in the group of mothers who were HBV- DNA positive in both serum and PBMC than that in the group of mothers who were serum HBV-DNA positive only( P <0.01); and it was markedly higher in the group of mothers who were PBMC HBV- DNA positive than that in group of mothers who were HBV-DNA negative in PBMCs( P <0.01). The positive rate of HBV-DNA in PBMCs of newborns was significantly higher in the group of newborns who were HBV-DNA positive in serum than that in the group of newborns who were HBV-DNA negative in serum( P <0.05).Conclusions The positive rate of HBV-DNA in PBMCs from newborns of HBsAg positive pregnant women was 30.0% (15/30). It was related to HBV viremia level and HBV-DNA status in PBMCs of mothers and newborns. Detection of HBV-DNA in PBMCs may be an important supplementary method to determine intrauterine HBV infection, and can predict the response to HBV vaccine.