HBsAg阳性孕妇胎盘组织HBV cccDNA检测及临床意义研究

发布时间：2018-01-15 14:23

本文关键词：HBsAg阳性孕妇胎盘组织HBV cccDNA检测及临床意义研究　出处：《桂林医学院》2015年硕士论文　论文类型：学位论文

【摘要】：目的:乙型肝炎病毒(Hepatitis B Virus,HBV)主要通过母婴垂直传播引起,其中宫内感染是最主要的传播途径,是造成免疫预防失败的主要原因。本实验将通过一种新型的原位检测方法,检测HBsAg阳性孕妇胎盘组织中HBVcccDNA存在情况,进一步探讨其意义及乙型肝炎病毒母婴传播机制,研究其与宫内感染发生的关系。方法:以52例北京302医院妇产中心进行产检并分娩的HBsAg阳性的孕妇产妇胎盘组织作为实验组,5例乙型肝炎病毒五项标志物均为阴性的健康孕妇的胎盘组织作为阴性对照组,同时以HBsAg与HBcAg均阳性的肝癌肝穿样本作为阳性对照组。收集孕妇及其新生儿临床资料,胎儿娩出后30min内于脐带中央切取2.0cm×2.0cm×1.5cm大小的胎盘组织,立即置于10%中性福尔马林中固定,经梯度酒精脱水、石蜡包埋、4.5μm连续切片后使用。Envision免疫组织化学染色方法检测石蜡包埋胎盘组织各类细胞中HBsAg、HBcAg表达情况;原位聚合酶链反应方法(in situ polymerase chain reaction,IS-PCR)检测胎盘组织中HBV cccDNA表达情况。每次实验均设立阴性对照组与阳性对照组。以新生儿出生后24h内股静脉血中HBsAg、HBeAg或HBV DNA任一项为阳性作为新生儿发生宫内感染的标志。结果:1.52例HBsAg阳性孕妇中,共有21例新生儿发生宫内感染,总的感染率为40.38%(21/52)。大三阳组孕妇新生儿HBV感染率为89.47%(17/19),高于小三阳组孕妇12.12%(4/33),P0.05,差异有统计学意义。HBV DNA高载量组(104~109IU/ml)与低载量组(102~104 IU/ml)新生儿HBV感染率分别为77.78%(14/18)、25.00%(3/12),P=0.014,差异有统计学意义。2.免疫组织化学染色实验组胎盘组织HBs Ag与HBcAg没有阳性信号,阳性对照组HBsAg与HBcAg均为阳性,阴性对照未检出阳性信号。3.原位聚合酶链反应在HBs Ag阳性孕妇胎盘组织中未检出HBV cccDNA阳性信号,阳性对照组HBV cccDNA检测有阳性信号,阴性对照组未检测到HBV ccc DNA阳性信号。结论:胎盘组织有可能不是发生HBV宫内感染的主要途径,可能通过其他途径发生感染。宫内感染的高危因素有待于大样本量的实验证实。
[Abstract]:Objective: hepatitis B virus (Hepatitis B, Virus, HBV) mainly through vertical transmission caused by intrauterine infection, which is the main route of transmission, is a major cause of immunization failure. This experiment will be through a novel in situ detection methods, HBVcccDNA detection of HBsAg positive placenta in the presence of further study the significance of maternal transmission of hepatitis B virus and study its mechanism, and the relationship between the occurrence of intrauterine infection. Methods: 52 cases of No.302 Hospital of Beijing Maternity Center for production inspection and delivery of the placenta HBsAg positive pregnant women as the experimental group, 5 cases of hepatitis B virus markers were five placentas of healthy pregnant women as negative the negative control group, while HBsAg and HBcAg were positive in liver samples as positive control group. Clinical data of pregnant women and their newborns, 30min after delivery of fetus The placenta is 2.0cm * 2.0cm * 1.5cm size in the central cord cut, immediately placed in 10% neutral formalin, after gradient alcohol dehydration, paraffin embedding, 4.5 m serial sections after using.Envision immunohistochemical staining method to detect the paraffin embedded placental tissue of various cell HBsAg, HBcAg expression; in situ polymerase chain (in situ polymerase chain reaction method, reaction, IS-PCR) expression of HBV cccDNA detection in placenta of each experiment were set up. The negative control group and positive control group. The newborn within 24h after femoral venous blood HBsAg, HBeAg or HBV DNA for any positive signs as neonates of intrauterine infection. Results: 1.52 cases HBsAg positive pregnant women, a total of 21 cases of neonatal intrauterine infection, the total infection rate was 40.38% (21/52). 3 group of neonatal HBV infection rate was 89.47% (17/ 19), higher than that of small Sanyang Group 12.12% (4/33), P0.05, there was a significant difference between the.HBV DNA group (104~109IU/ml) with high load and low load group (102~104 IU/ml) neonatal HBV infection rate was 77.78% (14/18), 25% (3/12), P=0.014, the difference has statistical significance of immunohistochemical.2. staining in experimental group placenta HBs Ag and HBcAg have no positive signal, positive control group HBsAg and HBcAg were all positive and negative controls were detected.3. positive signals in situ polymerase chain reaction cccDNA HBV positive signals were not detected in HBs Ag positive placenta, positive control group HBV cccDNA detected positive signal, the negative control group, positive signals were not detected by HBV CCC DNA. Conclusion: the placenta may not be the main route of HBV intrauterine infection, may be infected by other means. The risk factors of intrauterine infection remains to be confirmed by large sample experiments.

【学位授予单位】：桂林医学院
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R714.251

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