父婴传播的乙型肝炎病毒C基因变异研究  

HBV C gene mutation in the transmission from father to infant

作　　者：王珊珊[1] 李珉珉 彭桂福[1] 李文玲[3] 金惠玲[4] 肖红[1] 曾年华[1] 王志斌[1] 黄佳亮[1] 苏建新[1] 

机构地区：[1]广州军区军事医学研究所流行病学研究室,510507 [2]暨南大学医学院附属华侨医院检验科 [3]广州军区总医院妇产科 [4]陕西省西安医科大学第二附属医院妇产科

出　　处：《中华儿科杂志》2003年第11期845-848,共4页Chinese Journal of Pediatrics

基　　金：广东省卫生厅资助课题 (A2 0 0 15 2 5 )

摘　　要：目的　探讨乙型肝炎病毒 (HBV)父婴传播的可能性。方法　选择父亲为HBV携带者而母亲无任何HBV感染标志的新生儿为研究对象 ,检测新生儿的HBV感染标志 ,比较父亲与子代所携HBVC基因nt 2 0 2 2～ 2 30 1段序列同源性。结果　 16对父亲与新生儿所携HBVC基因同源性在99 %～ 10 0 % ,检出 2 0 2 9、2 0 34、2 0 4 4、2 0 5 9、2 0 78、2 0 95、2 10 4、2 15 4、2 16 1、2 16 9、2 189、2 2 0 1、2 2 33、2 2 5 1、2 2 84、2 2 88、2 2 93位核苷酸变异 ,其中 2 189、2 2 88位核苷酸变异为表型变异。 2 189位点A变C致使C区第 97位氨基酸由异亮氨酸变为亮氨酸 ,2 2 88位点C变A使C区第 130位氨基酸由脯氨酸变为苏氨酸 ,其他位点为无表型变异。 6对 (37 5 % )父亲与新生儿存在上述表型变异。Objective Hepatitis B virus (HBV) DNA was detected from infants whose mothers were negative for all HBV markers and the fathers were HBV carrier, the homology of HBV sequence of fathers and fetus was high, and HBV mutations concentrated on some points, and the transmission of HBV from father to fetus was also identified in some reports. The present study aimed to study HBV transmission from father to infant. Methods The study enrolled 16 pairs of fathers who were HBV carriers and infants whose mothers were negative for HBV markers. The infants had evidences for intrauterine HBV infection. The five HBV serum markers HBsAg, HBeAg, anti-HBe, anti-HBs, and anti-HBc were detected with ELISA. The positive results for HBsAg and/or HBeAg were regarded as markers of HBV infection. Amplification of HBV DNA was done using a nested PCR method. The first amplification was carried out using primer C1(nt 2394-2370), and primer C3(nt 1730-1754). The second amplification was carried out using primer C2(nt 1955-1974) and primer C6(nt 2348-2330). Both primers were designed to amplify the part of sequence coding for the hepatitis B C antigen. The size of the amplified fragment obtained by the nested PCR was expected to be 394 bp. The PCR products were electrophoresed on 1.5% agarose gels, which were then stained with ethidium bromide and observed with ultraviolet transillumination. When 394 bp specific band was detectable, the sample was designated positive. Then the positive samples were identified by dot blot. The second PCR products were extracted by phenol-chloroform and 70% ethanol precipitation, then resuspended in TE buffer (pH8.0), and used as the template for cloning. The template was connected into pGEM-T vector by ligase. The ligated products were cloned into fresh competent JM109 cells, and incubated for 90 minutes at 37℃ on roller drum. Finally several dilutions were plated on plates containing ampicillin, X-Gal and IPTG, and incubated at 37℃ overnight. The white colony on plates was used for identification by

关 键 词：父婴传播 乙型肝炎病毒 C基因 基因变异 序列分析 垂直疾病传播 

分 类 号：R181.3[医药卫生—流行病学]