宁夏地区HBV基因分型、耐药突变及HLA-DRB1基因多态性与乙型肝炎进展的相关性研究

发布时间：2018-06-01 09:39

  本文选题：乙型肝炎病毒 + HBV-DNA　； 参考：《宁夏医科大学》2014年硕士论文

【摘要】：目的调查宁夏地区慢性乙型肝炎（Chronic hepatitis B，CHB）患者的ALT、HBV-DNA载量与血清标志物水平之间的关联性，探讨HBV基因型、核苷（酸）类似物（nucleostide analogues，NAs）耐药突变模式与HBV感染后慢性化、重症化的关系，研究HLA-DRB1基因多态性与乙肝相关性疾病的关系。 方法随机选取CHB患者283例，应用化学发光微粒子免疫分析法定量检测HBV血清学标志物和荧光定量PCR检测HBV-DNA载量，并对结果进行相关性分析。从中选取134例住院患者的血清标本应用基因型特异性多对引物巢式PCR法进行HBV基因分型，再使用直接测序法对分型结果进行验证并检测HBV的耐药突变，，并针对不同的基因型进行性别、年龄、肝功能、HBV-DNA载量及HBeAg水平的相关性分析，以及对不同耐药突变模式CHB患者之间的临床参数进行分析比较。另选取87例CHB患者、78例肝硬化患者、31例肝癌患者和50例健康对照者作为研究对象，采用序列特异性引物-聚合酶链式反应（polymerase chain reaction-sequence specificprimers PCR-SSP）方法对他们的HLA-DRB1等位点进行基因分析研究。 结果（1）HBeAg阳性患者较阴性患者的HBeAb水平和HBV-DNA载量均有显著性差异（P0.001，P0.001）；女性的HBeAg和HBeAb水平均较男性高（P0.05，P0.01）；30-50岁组的CHB患者的HBV-DNA、HBeAg、HBeAb与30岁组、30-50岁组比较有显著性差异（P0.01，P0.001，P0.01）；CHB患者中HBV DNA载量与HBeAg呈正相关（r=0.451，P0.001），与HBeAb呈正相关（r=0.434，P0.001)；ALT与HBsAg呈正相关（r=0.131，P0.05）。 （2）134例HBV感染者的血清标本中，B基因型11例（8.2%），其中男10例，女1例；C基因型123例（91.8%），男87例，女36例。B型和C型之间的HBV-DNA和ALT水平有显著差异（Z=3.23，P0.05；Z=0.19，P0.05）。全部标本中有28例发生不同位点变异，变异率为20.9%，以单位点rtS213T突变为主，约占25.0%。 （3）与健康对照组相比，乙肝后肝硬化患者的DRB1*07、DRB1*12等位基因表达频率明显高于健康对照组，差异显著（P0.05，OR=2.237，95%CI为1.689～2.961；P0.05，OR=2.317，95%CI为1.707～3.143）；汉族乙肝相关性疾病患者的DRB1*04、DRB1*13、DRB1*15等位基因表达频率与汉族健康对照者的差异显著（P0.05，OR=0.478，95%CI为0.367～0.623；P0.05，OR=0.462，95%CI为0.355～0.602；P0.05，OR=2.292，95%为1.599～3.283）。 结论（1）宁夏地区的慢性乙型肝炎患者的HBV-DNA载量与HBeAg呈正相关，两者之间具有良好的一致性；但ALT水平与HBsAg水平呈正相关；女性患者的HBeAg浓度较男性高，其余指标无显著性差异。慢性乙型肝炎患者以30-50岁年龄阶段的居多，且此组患者的HBV-DNA、HBeAg、HBeAb与其余年龄段组比较有显著性差异。 （2）宁夏地区慢性乙型肝炎患者的基因型包括B型、C型，其中以C基因型为主，其次为B基因型；HBV基因型与HBV-DNA、ALT水平有关联，和本研究中其他指标比较无显著差异；CHB患者主要对拉米夫定（LAM）与阿德福韦酯（ADV）耐药，出现以单位点rtS213T为主的突变，并出现了混合位点突变。 （3）HLA-DRB1*07、DRB1*12可能是宁夏地区乙肝患者发生肝硬化的易感基因；HLA-DRBI*04和DRBI*l3可能是宁夏地区汉族人群的抗性基因；携带DRB1*15的汉族人群更容易发生乙肝相关性疾病。
[Abstract]:Objective To investigate the relationship between HBV - DNA load and serum marker level in patients with chronic hepatitis B ( HBV ) in Ningxia , and explore the relationship between HBV genotype , nucleoside ( acid ) analog ( NAs ) resistance mutation pattern and chronic and severe HBV infection , and study the relationship between HLA - DRB1 gene polymorphism and hepatitis B - related diseases .

Methods A total of 283 patients were randomly selected , and the HBV - DNA content was determined quantitatively by using the method of chemiluminescence , and the correlation of HBV - DNA level and HBV - DNA level was analyzed by direct sequencing .

Results ( 1 ) There was significant difference between HBeAg - positive patients and HBeAg - positive patients ( P0.001 , P0.001 ) .
The levels of HBeAg and HBeAg in women were higher than those in males ( P0.05 , P0.01 ) .
HBV - DNA , HBeAg , HBeAg , HBeAg , HBeAg , HBeAg , HBeAg , HBeAg , HBeAg and HBeAg were significantly different between 30 - 50 years old and 30 - 50 years old ( P0.01 , P0.001 , P0.01 ) .
The amount of HBV DNA was positively correlated with HBeAg ( r = 0.451 , P0.001 ) and positive correlation with HBeAg ( r = 0.434 , P0.001 ) .
ALT was positively correlated with HBsAg ( r = 0.131 , P0.05 ) .

( 2 ) Of 134 patients with HBV infection , 11 cases ( 8.2 % ) had genotype B , 10 were male and 1 female ;
There were 123 cases of genotype C ( 91.8 % ) , 87 males and 36 females . There was significant difference in HBV - DNA and ALT levels between genotype B and C ( Z = 3.23 , P0.05 ) .
Z=0.19,P0.05). In all the specimens , 28 cases had different site variation , and the mutation rate was 20 . 9 % . rtS213T mutation was dominant at the unit point , accounting for 25.0 % .

( 3 ) The frequency of DRB1 * 07 and DRB1 * 12 allele in patients with cirrhosis after hepatitis B was significantly higher than that in healthy control group ( P0.05 , OR = 2.237 , 95 % CI was 1.689 ~ 2.961 ) .
P0.05,OR=2.317,95%CI涓

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