HBsAg阳性孕妇早期流产绒毛组织中HBV标志物检测及风险因素研究  

Risk factor and hepatitis B virus makers during the first trimester in villi tissues of HBsAg-positive pregnant women

作　　者：彭婷婷 王淼[2] 陈凤 喻宏 杨敏 黄华鑫 崔璐[2] 蔡秋娥[2] 王强[2] 苟继周[2] 孙炎[2] 陈楚明 刘映霞[2] Peng Tingting;Wang Miao;Chen Feng;Yu Hong;Yang Min;Huang Huaxin;Cui Lu;Cai Qiu’e;Wang Qiang;Gou Jizhou;Sun Yan;Chen Chuming;Liu Yingxia(University of South China, Hengyang 421000, China;State Key Discipline of Infectious Diseases, Department of Infectious Diseases, Shenzhen Third People′s Hospital, Shenzhen 518112, China)

机构地区：[1]南华大学,衡阳421000 [2]深圳市第三人民医院,感染病国家重点专科,518112

出　　处：《中华实验和临床病毒学杂志》2019年第2期182-186,共5页Chinese Journal of Experimental and Clinical Virology

基　　金：国家自然科学基金(81570552,81873573,81800525);"十三五"艾滋病和病毒性肝炎等重大传染病防治重大专项(2108ZX10711001).

摘　　要：目的通过检测乙型肝炎表面抗原(hepatitis B surface antigen,HBsAg)阳性孕妇早期流产的绒毛组织中与乙型肝炎病毒(hepatitis B virus,HBV)感染相关的指标,评估妊娠早期宫内感染的发生情况及可能的影响因素。方法以45例HBsAg阳性孕妇早期流产的绒毛组织和血液标本为研究对象,应用荧光定量qRT-PCR方法,对HBV DNA进行测定,并与血液中HBV DNA的检测结果进行比较分析;应用免疫组化技术对绒毛组织中Glial Cell Missing-1(GCM1)、HBsAg、HBcAg进行检测。结果qRT-PCR检测结果显示,45份绒毛组织中有14份样本HBV DNA呈阳性,阳性率为31.11%;血液检查结果中有24份样本呈阳性,阳性率为53.33%,进一步统计分析的结果表明血液和绒毛组织的HBV DNA阳性率间无显著性差异(χ^2=4.555,P=0.054)。其中,有12份绒毛组织样本与血液样本阳性结果相符合,且这部分孕妇外周血的HBsAg、HBeAg、HBeAb和HBV DNA水平均显著高于其他孕妇(P值分别为0.007、0.004、0.000、0.000)。多因素logistic回归分析表明血液HBV DNA大于10^6 IU/ml与绒毛组织HBV DNA阳性独立相关,且这部分孕妇的HBsAg、HBeAg、绒毛组织HBV DNA阳性率均显著高于其他孕妇(P值均为0.000)。免疫组化结果表明,45份绒毛组织的胞核均表达其特异性抗体GCM1;有9份绒毛组织的胞浆表达HBsAg;只有1份绒毛组织的胞核表达HBcAg。结论HBsAg阳性孕妇早期流产的绒毛组织中可检测到HBV DNA、HBsAg的表达,提示胎儿,尤其是高病毒载量母亲所怀的胎儿早期存在发生宫内感染的可能。Objective To determine whether intrauterine infection with hepatitis B virus (HBV) occurs in early pregnancy and to characterize associated virulence factors. Methods Villi tissues and blood samples of 45 HBV surface antigen (HBsAg)-positive pregnant women were collected during the first trimester and HBV DNA loads were quantified by quantitative reverse transcription polymerase chain reaction (qRT-PCR). The expression of GCM1, HBsAg and hepatitis B core antigen (HBcAg) in villi tissues were detected by immunohistochemical method. Results Data from qRT-PCR showed that HBV DNA was detected in 14 of 45 villi tissues (positive rate of 31.11%), and 24 of 45 blood samples (positive rate of 53.33%), further statistical analysis showed that the positive rates of HBV DNA between blood samples and villi tissues were not significantly different (χ^2=4.555, P=0.054). Among them, 12 samples were consistently positive between the villi and blood specimens, and HBsAg, HBeAg, HBeAb, HBV DNA from peripheral blood in these pregnant women were significantly higher than those of the other women (P value was 0.007, 0.004, 0.000, and 0.000 respectively). The multivariate logistic regression analysis showed that blood HBV DNA greater than 10^6 IU/ml was independently associated with HBV DNA positive in villi, and the HBsAg, HBeAg, villi tissues HBV DNA positive rates of these pregnant women were significantly higher than those of the other pregnant women (all P value were 0.000). Immunohistochemistry results showed that all 45 cases were positive for GCM1 expression in the cell nucleus. Nine cases also had HBsAg expression in the cytoplasm. Only one case was found to express HBV core antigen (HBcAg) in the nucleus. Conclusions HBV DNA and HBsAg can be detected from villi tissues harvested during the first trimester in HBsAg-positive pregnant women, and the results suggest an early occurrence of intrauterine infection of fetuses with high HBV levels.

关 键 词：肝炎病毒 乙型 绒毛组织 QRT-PCR 免疫组化 宫内感染 

分 类 号：R714.21[医药卫生—妇产科学]