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目的探讨应用套式PCR方法检测乙型肝炎表面抗原(HBsAg)及乙型肝炎e抗原(HBeAg)阴性孕妇乙型肝炎病毒(HBV)宫内感染的状况。方法选择HBsAg与HBeAg阴性,其他HBV血清标志物阳性孕妇及其新生儿24例作为病例组,同期HBV血清标志物全部阴性孕妇及其新生儿16例作为对照组。采用套式PCR方法检测两组孕妇及其新生儿的血清及外周血单个核细胞(PBMC)中HBVDNA。结果(1)病例组24例孕妇中,血清HBVDNA阳性8例,阳性率为33%;PBMC中HBVDNA阳性10例,阳性率为42%。其中血清与PBMC均阳性3例,总阳性率为63%(15/24)。(2)病例组24个新生儿中,血清HBVDNA阳性3例,阳性率为13%,PBMC中HBVDNA阳性6例,阳性率为25%。其中血清与PBMC均阳性1例,宫内感染率为33%(8/24)。(3)病例组24例孕妇中,血清阴性而PBMC阳性共7例,其新生儿4例发生宫内感染,感染率为4/7。(4)对照组16例孕妇及其新生儿血清及PBMC中HBVDNA全部阴性。结论HBsAg及HBeAg阴性孕妇也可发生HBV宫内感染,采用灵敏度高的套式PCR方法检测孕妇及其新生儿血清及PBMC中HBVDNA,对诊断HBV宫内感染具有重要临床意义。
Objective To investigate the detection of hepatitis B virus (HBV) intrauterine infection in hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) negative pregnant women by nested PCR. Methods 24 pregnant women and their newborns with negative HBsAg and HBeAg were selected as the case group and 16 pregnant women and their newborns with negative HBV seropositive for the same period as the control group. The nested PCR method was used to detect HBVDNA in serum and peripheral blood mononuclear cells (PBMCs) of two groups of pregnant women and their newborns. Results (1) Serum HBVDNA was positive in 8 out of 24 pregnant women in the case group, the positive rate was 33%. The positive rate of HBVDNA in PBMC was 10% (42%). Serum and PBMC were positive in 3 cases, the total positive rate was 63% (15/24). (2) Of the 24 newborns in the case group, 3 were positive for serum HBVDNA, the positive rate was 13%, and 6 were HBVDNA positive in PBMC. The positive rate was 25%. Serum and PBMC were positive in 1 case, intrauterine infection rate was 33% (8/24). (3) Of the 24 pregnant women in the case group, 7 were seronegative and PBMC positive, with 4 neonates with intrauterine infection, the infection rate was 4/7. (4) HBVDNA in serum and PBMC of 16 pregnant women and their newborns in the control group were all negative. Conclusions HBsAg and HBeAg-negative pregnant women may also develop intrauterine infection of HBV. Using high sensitivity nested PCR to detect HBVDNA in serum and PBMC of pregnant women and their newborns, it is of great clinical significance to diagnose HBV intrauterine infection.