慢性肝炎和肝癌病人血清中乙型肝炎病毒DNA的检测

为了了解慢性肝炎和肝癌病人患者体内乙型肝炎病毒（ＨＢＶ）复制与ＨＢＶ血清标志之间的关系，用酶联免疫吸附实验（ＥＬＩＳＡ）、聚合酶链反应（ＰＣＲ）及斑点杂交方法对６１例慢性肝炎和４７例肝癌患者的ＨＢＶ表面抗原（ＨＢｓＡｇ）、相关ｅ抗原（ＨＢｅＡｇ）、表面抗体（抗－ＨＢｓ）、核心抗体（抗－ＨＢｃ）、相关ｅ抗体（抗－ＨＢｅ）进行了检测。结果表明：ＨＢＶＤＮＡ在ＨＢｓＡｇ、ＨＢｅＡｇ、／抗－ＨＢｃ阳性的慢性肝炎和肝癌患者血清中的检出率分别为９０．５０％和５０．００％；在ＨＢｓＡｇ／抗－ＨＢｅ、抗－ＨＢｃ阳性者的检出率分别为４５．４０％和７．１４％；在ＨＢｓＡｇ阳性、ＨＢｅＡｇ阴性／抗－ＨＢｅ阴性者中的检出率分别为６０．００％和４０．００％；ＨＢｓＡｇ阴性、／抗－ＨＢｃ阳性或／抗－ＨＢｅ阳性或／抗－ＨＢｓ阳性者中的检出率分别为２０．００％和２２．２２％；在血清学指标全阴性时，慢性肝炎和肝癌患者血清中ＨＢＶＤＮＡ的检出率均为０。实验提示：无论是肝炎或肝癌，在ＨＢｓＡｇ、ＨＢｅＡｇ同时阳性时，ＨＢＶ复制最为活跃；在单独ＨＢｓＡｇ阳性时，ＨＢＶ有一定程度的复制；ＨＢＶ复制在肝癌细胞中受到一定程度的抑制。     

血清乙型肝炎病毒前S1抗原检测及其与病毒复制的关系

用抗Ｓ和抗前Ｓ１单抗的双抗体夹心ＥＬＩＳＡ法检测１５０例慢性乙型肝炎患者、乙型肝炎病毒表面抗原（ＨＢｓＡｇ）携带者和健康人血清中的ＨＢＶ前Ｓ１抗原，其结果和ＨＢＶＤＮＡ聚合酶链反应（ＰＣＲ）、乙型肝炎血清标志的检测结果进行比较。结果表明：前Ｓ１抗原在乙型肝炎病毒ｅ抗原（ＨＢｅＡｇ）阳性组中的检出率和相对滴度显著高于ＨＢｅＡｇ阴性组（Ｐ＜０．０１）；在ＨＢｅＡｇ阴性组中，抗－ＨＢｅ阴性人群前Ｓ１抗原的检出率和相对滴度也显著高于抗－ＨＢｅ阳性人群（Ｐ＜０．０１）。前Ｓ１抗原和ＨＢＶＤＮＡ检测结果的符合率达８０％，两者检出率的相关系数ｒ＝０．９８２６（Ｐ＜０．０１）。结论：血清前Ｓ１抗原和乙型肝炎病毒的存在关系密切。     

受体导向药物L-HSA-Ara-AMP对乙型肝炎病毒抑制作用的临床初探

目的为初步试探受体导向药物Ｌ－ＨＳＡ－Ａｒａ－ＡＭＰ的抗病毒效果。方法应用５天疗法治疗慢性乙型肝炎１０例，并以Ａｒａ－ＡＭＰ为对照。结果治疗组ＨＢｓＡｇ无变化，１０例ＨＢｅＡｇ阳性者阴转３例，滴度下降６例，４例抗－ＨＢｃＩｇＭ阳性均阴转，８例ＨＢＶＤＮＡ阳性阴转４例，浓度下降４例，未发现任何毒副反应。结论显示该药对ＨＢＶ复制指标的近期抑制效果与Ａｒａ－ＡＭＰ相近或略优，但日剂量仅为后者的１／７。     

慢性乙型肝炎重叠HGV感染对HBV复制的影响

目的观察慢性乙型肝炎重叠ＨＧＶ感染对ＨＢＶ复制的影响．方法采用逆转录聚合酶链反应（ＲＴ－ＰＣＲ）检测患者血清ＨＢＶ－ＤＮＡ．ＨＢＶ－ＤＮＡ定量采用荧光信号引物能量转换法．以单纯慢性乙型肝炎患者作对照研究．结果２３例ＨＧＶ重叠感染的慢性乙型肝炎患者 ＨＢＶ－ＤＮＡ、ＨＢｅＡｇ和抗－ＨＢｅ的阳性率分别为５６．５％、１７．４％和６０．９％而３４例单纯慢性乙型肝炎患者三项指标的阳性率分别为９１．２％、８５．３％和１１．８％．定量分析显示,前组ＨＢＶ－ＤＮＡ量为２．０２、后组为４．１２．经统计学处理,两组各项指标均有显著性差异．结论慢性乙型肝炎重叠ＨＧＶ感染可能会抑制ＨＢＶ的复制．     

不同慢性肝病患者血清中乙型肝炎病毒DNA定量检测及其临床意义

目的探讨乙型肝炎病毒ＤＮＡ含量的临床意义。了解乙型肝炎病毒（ＨＢＶ）免疫标志不同状态的慢性肝病患者血清ＨＢＶＤＮＡ浓度及其临床意义。方法应用建立的竞争性聚合酶链反应（ＰＣＲ）方法定量检测慢性肝炎（ＣＨ）５１例、肝硬化（ＬＣ）３６例、原发性肝癌（ＰＨＣ）３８例的血清ＨＢＶＤＮＡ浓度。结果ＨＢＶＤＮＡ阳性的ＣＨ患者血清ＨＢＶＤＮＡ浓度为４．３６ｌｏｇ１０ＨＢＶＤＮＡ拷贝５０μｌ（下同），ＬＣ为４．５５，ＰＨＣ为４．４３，三组间无显著性差异（Ｐ＞０．０５）；血清ＨＢＶ五项免疫标志均阴性或抗－ＨＢｓ阳性的慢性肝病患者中，有３７．５％患者存在低水平ＨＢＶ复制；ＨＢｅＡｇ阳性患者的ＨＢＶＤＮＡ浓度总体上明显高于抗－ＨＢｅ阳性组，但其中部分患者的ＨＢＶＤＮＡ浓度也很高。结论提示ＨＢＶ的复制状态与慢性肝病的病期无明显关系；在抗－ＨＢｅ阳性的患者中存在个体差异，故不能仅依据抗－ＨＢｅ阳转来判断ＨＢＶ复制减少或停止。     

肝细胞内HBcAg和HBsAg分布的演变特点

本文报道用免疫组化（ＡＢＣ）法检测２９例慢性乙型肝炎患者肝活检组织中ＨＢｃＡｇ和ＨＢｓＡｇ分布的动态演变，结合血清ＨＢｅＡｇ和ＨＢＶ ＤＮＡ的演变探讨其病毒学意义。ＨＢｃＡｇ、ＨＢｅＡｇ和ＨＢＶ ＤＮＡ全阳性反映ＨＢＶ高复制相，全阴性为非复制相，不一致为两相之间的过渡型（低复制相）。肝细胞内ＨＢｃＡｇ由核浆型为主演变为核型或浆型为主乃至阴性意味着ＨＢＶ从高复制相转入低复制相乃至非复制相，同时伴有血     

受体导向药物L—HSA—Ara—AMP对乙型肝炎病毒抑制作？…

目的 为初步试探受体导向药物Ｌ－ＨＳＡ－Ａｒａ－ＡＭＰ的抗病毒效果。方法 应用５天疗法治疗慢性乙型肝炎１０例，并以Ａｒａ－ＡＭＰ为对照。结果 治疗组ＨＢｓＡｇ无变化，１０例ＨＢｓＡｇ阳性者阴转３例，滴度下降６例，４例抗－ＨＢｃ ＩｇＭ阳性均转阴，８例ＨＢＶ ＤＮＡ阳性阴转４例，浓度下降４例，未发现任何毒副反应。结论 显示该药对ＨＢＶ复制指标的近期抑制效果与Ａｒａ－ＡＭＰ相近或略优，但日剂量仅为后者     

HBcAg/HBeAg对慢性乙型肝炎患者PBMC中Th1/Th2类细胞应答的影响

目的 探讨ＨＢｃＡｇ／ＨＢｅＡｇ对慢性乙型肝炎患者ＰＢＭＣ中Ｔｈ１／Ｔｈ２类细胞应答的影响。方法 用套式ＰＣＲ法检测６４便慢性ＨＢＶ感染者ＰＢＭＣ中ＨＶＢ ＤＮＡ;分别用ＰＨＡ、ＨＢｃＡｇ和ＨＢｅＡｇ体外培养;ＥＬＩＳＡ法检测ＰＢＭＣ产生Ｔｈ１类细胞因子（ＩＬ－２、ＩＦＮ－γ）和Ｔｈ２类细胞因子（ＩＬ－４、ＩＬ－１０）的含量。结果 表明ＨＢＶ ＤＮＡ阳性组和阴性组相比,无论是在ＰＨＡ还是在ＨＢｃＡ     

HBV DNA含量与HBV标志物表现形式的初步研究

1　资料与方法ＨＢＶ感染者 6 6例 ,诊断按 1995年北京会议标准 ,其ＨＢＶＭ表现形式为 :ＨＢｓＡｇ、ＨＢｅＡｇ、抗ＨＢｃ均阳性 (大三阳 )4 2例 ,ＨＢｓＡｇ、抗ＨＢｅ、抗ＨＢｃ阳性 (小三阳 ) 14例 ,仅抗ＨＢｃ阳性 10例。抗病毒治疗方法 :无环鸟苷 (湖北潜江制药厂 )每日 2 0 /ｋｇ体重、分两次静滴、15ｄ后给予干扰素 (深圳科兴公司 ) 30 0万Ｕ/支、隔日 1支 ,共 5 0支。ＨＢＶＭ检测采用ＥＩＡ法 ,斑点杂交试剂为北京医科大学肝病研究所产品 ,操作按说明书进行。ＨＢＶＤＮＡ定量测定采用信号引物能量转移ＰＣＲ法 (ＱＰＣＲ) ,统计…     

154例HBV血清学标志物阳性PCR检测结果分析

１５４例ＨＢＶ血清学标志物阳性ＰＣＲ检测结果分析黄华健邵阳市中心医院检验科为了解ＨＢＶ—ＤＮＡ与乙型肝炎５种血清学标志物——即乙肝二对半（ＨＢｓＡｇ、Ａｎｔｉ—ＨＢｓ，ＨＢｅＡｇ，Ａｎｔｉ—ＨＢｅ，Ａｎｔｉ—ＨＢｃ）之间关系，本文对１５４例乙肝二对半...     

乙型肝炎病毒 C 基因启动子基因变异与免疫学标志及DNA含量的临床研究

目的 研究乙型肝炎病毒（HBV）C基因启动子（Basic core promoter,BCP）基因变异与HBV感染者免疫学标志及HBV DNA含量的临床关系。方法 采用聚合酶链（PCR）微板核酸分子杂交及荧光定量PCR检测技术,对246例HBV感染者进行HBV BCP基因变异及HBV DNA含量测定。结果 在HBsAg/HBeAg/HBcAB,HBsAg/HBeAg/HBcAb及HBsAg/HBcAb阳性的HBV感染者中,HBV DNA的阳性率分别为97.5%(48/49),22.9%(25/109),31.1%(14/45);HBV BCP基因变异率分别为59.1%(29/49),11%(12/109),13.3%(6/45);HBV BCP基因变异组DNA含量均≥10^6cps/ml,明显高于非BCP基因变异组。肝硬化病人BCP基因变异明显高于其他组。结论 e抗原免疫学标志的转换仅对部分感染者预示着病毒的免疫清除和静息,单独依靠乙型肝炎免疫学标志并不能确切提示HBV的复制状态、病变程度及愈后。     

奉贤地区HBsAg阴性的HBV自然感染母亲对新生儿影响的研究

目的　为揭示HBsAg阴性的乙型肝炎病毒 (HBV)自然感染孕妇的宫内感染及其危险因素。方法　采用多聚酶链反应 (PCR)技术结合酶联免疫吸附法 (ELISA) ,对奉贤地区 131例HBsAg阴性的HBV自然感染孕妇外周血 ,及其分娩后的脐带血进行HBV血清学标志物 (HBVM)和HBVDNA检测。结果　HBsAg阴性的HBV自然感染孕妇宫内的感染率 (除外单一抗 -HBs阳性 )为 5 2 6 7% ;脐血中不同HBVM组合的HBVDNA检出率依次为 :抗 -HBe( )、抗 -HBc( ) >抗 -HBs( )、抗 -HBe( )、抗 -HBc( ) >抗 -HBs( )、抗 -HBe( ) >抗 -HBs( )、抗 -HBc( ) >抗 -HBs( ) ;脐血HBVDNA总检出率为 16 79%。结论　HBsAg阴性的HBV自然感染孕妇也可能发生宫内感染。提议HBsAg阴性的HBV自然感染孕妇和新生儿有进行自动和被动免疫接种的必要性     

Molecular analysis of the hepatitis B virus presurface and surface gene in patients from eastern China with occult hepatitis B

This study was designed to detect and analyze mutations that occur within the presurface and surface (pre‐S/S) gene of HBV in patients with occult hepatitis B, and determine their relationship to that disorder. Among 254 HBsAg negative samples of blood collected in eastern China, 183 were positive for anti‐HBc alone, 61 were positive for anti‐HBe alone, and 10 samples were positive for HBeAg. Within this group, 15 samples were found to be HBV DNA positive by real‐time PCR and were designated Group I. A control group of 28 HBsAg positive samples were chosen at random from patients with chronic hepatitis B and designated Group II. The HBV pre‐S/S gene was amplified by PCR and subjected to sequencing analysis. Occult hepatitis B was found in 1.6% of the patients with anti‐HBc alone and in 3.3% of those with anti‐HBe alone. Occult hepatitis B also was found in all HBsAg negative but HBeAg positive samples. Sequencing analysis showed a significant correlation between point mutations within the “a” determinant and occult hepatitis B (P < 0.0001), and a close relationship between pre‐S deletion mutations and occult hepatitis B (P = 0.06). There were unique amino acid mutations at the G145 position other than G145R. The HBV DNA levels in patients with occult hepatitis B were significantly lower than those found in the control group. The “a” determinant mutations and pre‐S deletions may play important roles in occult hepatitis B by affecting the expression, synthesis and secretion of the S protein and by impeding viral release and replication. J. Med. Virol. 85: 979–986, 2013. © 2013 Wiley Periodicals, Inc.     

Reactivation of occult hepatitis B virus infection following cytotoxic lymphoma therapy in an anti‐HBc negative patient

Screening hepatitis B virus (HBV) surface antigen (HBsAg) and HBV core antibody (anti‐HBc) is recommended prior to cytotoxic or immunosuppressive therapy. This case describes an anti‐HBc negative, DNA positive occult HBV infection in a 71‐year‐old Caucasian male following rituximab‐based treatment for follicular lymphoma. Pre‐screening serology indicated negative HBsAg and anti‐HBc. However, following sequential treatment cycles the patient developed weak HBsAg with a low HBV DNA load (<1,000 IU/ml), but remained anti‐HBc negative. The DNA load peaked 5 months later (>1 × 10⁶ IU/ml) and he was subsequently treated with Tenofovir. Currently the patient remains anti‐HBc negative, and is anti‐HBe negative, anti‐HBs negative, HBeAg positive. No clinical or biochemical evidence of hepatitis has occurred. Sequencing and phylogenetic analysis identified the HBV genosubtype as D4, most probably acquired some years ago during a stay in Papua New Guinea, in spite of prior hepatitis B vaccination. Four amino acid substitutions were detected within the HBsAg loop yet none in the core protein. This case questions the dependability of anti‐HBc testing and highlights the role of HBV DNA testing prior to and throughout cytotoxic or immunosuppressive regimes. As this case exemplifies, vaccination protects against clinical infection but may not exclude seronegative occult infection with the possibility of reactivation. J. Med. Virol. 85:597–601, 2013. © 2013 Wiley Periodicals, Inc.     

山东省枣庄市乙型病毒性肝炎流行病学调查

目的　了解枣庄市人群中乙型肝炎的流行特征。方法　于 2 0 0 0年采用随机分层抽样 ,调查 312户家庭的 96 3人 ,以RIA法检测HBsAg、抗 HBs和抗 HBc。结果　HBsAg、抗 HBs、抗 HBc和HBV标化流行率分别为 7.0 8%、37.5 6 %、4 1.35 %和 4 4 .37%。HBsAg流行率男性高于女性 (P <0 .0 5 ) ,城区高于农村 (P <0 .0 1) ,在不同年龄及职业人群中差异无显著性 (P >0 .0 5 )。抗 HBs、抗 HBc和HBV感染率有随年龄增长而递增的趋势 (P <0 .0 1)。HBV总感染率男性高于女性 (P <0 .0 5 ) ,农村高于城市 (P <0 .0 5 )。结论　枣庄市人群HBV感染率较高 ,应积极采取预防和控制措施 ,减少发病。     

乙肝患者外膜蛋白血清学检测及对于判定HBV DNA复制的意义

目的探讨HBV感染者血清中PreS1-Ag、PreS2．Ag、大蛋白（LP）的检测意义及其对判定HBV复制的意义。方法应用酶联免疫吸附试验（ELISA）检测201例HBV感染血清的PreS1-Ag、PreS2-Ag、HBV-LP及HBVM，同时应用荧光定量PCR方法检测HBVDNA。结果PreS1-Ag、PreS2-Ag、LP、HBsAg阳性率差异有统计学意义，PreS2-Ag、LP检出阳性率均高于HBeAg；LP的检出阳性率与HBVDNA的检出阳性率相关性有统计学意义，且HBV DNA拷贝数的对数值与HBV-LP表达呈正相关。结论PreS1-Ag、PreS2-Ag、LP较准确的反映乙肝病毒的复制情况，是HBVM有益的必要补充；血清中HBV-LP的含量与HBVDNA的拷贝数具有较好的相关性。     

检测乙型肝炎病毒表面抗原大蛋白的临床意义

目的检测乙肝患者血清中乙肝两对半（HBVM）、乙型肝炎病毒表面抗原大蛋白（HBV—LP）以及HBVDNA，比较在不同乙肝两对半模式的患者中HBV．LP与HBVDNA的检出率，探讨乙型肝炎病毒表面抗原大蛋白（HBV—LP）用于乙肝患者临床诊断的意义。方法采用酶联免疫吸附实验（ELISA）检测HBV．LP和乙肝两对半，采用荧光定量PCR方法对患者HBVDNA进行检测。结果（1）相同乙肝模式患者血清中HBV．LP与HBVDNA检出率差异无统计学意义；（2）143例小三阳乙肝患者血清中HBV．LP与HBVDNA阳性率差异无统计学意义，二者的检出一致率为82．52％[（65＋53）／143]；（3）HBV—LP吸光度（A值）与HBVDNA呈正相关关系（r=0．983）。结论乙型肝炎病毒表面抗原大蛋白与HBVDNA有良好的正相关关系，在HBeAg阴性的乙肝患者中乙肝病毒表面抗原大蛋白（HBV—LP）与HBVDNA有较高检出一致率，HBV—LP用于临床反映乙肝病毒复制水平，特别是HBeAg阴性乙肝患者体内病毒复制情况有重要的意义。     

聚乙二醇干扰素α-2a联合基因重组酵母乙肝疫苗治疗HBeAg阳性CHB的疗效观察

目的 探讨聚乙二醇干扰素α-2a联合基因重组酵母乙肝疫苗治疗HBeAg阳性的慢性乙型肝炎患者疗效.方法 总75例HBeAg阳性慢性乙型肝炎纳入本研究,其中单用聚乙二醇干扰素α-2a治疗的45例（A组）;聚乙二醇干扰素α-2a联合基因重组酵母乙肝疫苗的HBeAg阳性慢性乙肝患者30例（B组）.对比分析两组在治疗0、24、48和72周时ALT、HBsAg水平、HBeAg血清转换率和HBV DNA阴转率的差异.结果 治疗前（0周）时两组患者的年龄、ALT、HBsAg和HBV DNA水平差异均无统计学意义（P＞0.05）,其中联合治疗组（B组）HBeAg水平明显高于对照组（A组）,差异具有统计学意义（P＜0.05）.第24周和48周时,两组患者的ALT、HBsAg水平、HBeAg血清学转换率和HBV DNA阴转率差异并无统计学意义（P＞0.05）.在治疗结束随访至72周时,A、B两组ALT、HBeAg血清转换率和HBsAg水平差异没有统计学意义（P＞0.05）,但B组HBV DNA阴转率高于A组,差异具有统计学意义（P=0.032）.结论 聚乙二醇干扰素α-2a联合基因重组乙肝疫苗治疗HBeAg阳性的慢性乙型肝炎患者可以提高48周治疗结束后72周时的HBV DNA阴转率,但是与HBeAg血清学转换和HBsAg水平降低无关.     

Hepatitis B virus DNA in patients with HBsAg in south western Nigeria

There are about 400 million people with chronic hepatitis B virus (HBV) infection worldwide with a potential of adverse sequelae including hepatocellular carcinoma. Recent data have shown that the level of HBV DNA in serum or plasma of an infected person probably reflects more accurately the replicative activity of the virus and therefore may serve as a better maker for management of the infection. This study was designed to determine the rate of detection of HBV DNA in blood samples of patients with HBsAg positive in Nigeria in comparison with the HBe and anti‐HBe used widely as serological markers of infectivity. Plasma samples from 105 patients with HBsAg positive were tested for the presence of HBeAg and anti‐HBe using a commercial enzyme‐linked immunosorbent assay while plasma HBV DNA was quantified using the COBAS Amplicor HBV Monitor assay. Of the 105 HBsAg samples, 17 (16.2%) and 85 (81%) were positive for HBeAg and anti‐HBe, respectively, while 8 (7.6%) were negative for both HBeAg and anti‐HBe. HBV DNA was detected in 86 (81.9%) of the samples, out of which 15 (18.1%) and 67 (80.7%) were positive for HBeAg and anti‐HBe, respectively. HBV DNA was detected in 78.4% of the HBeAg negative samples and in all the eight samples that were negative for both HBeAg and anti‐HBe. The implication of these findings in the management of patients with HBV infection is compelling. J. Med. Virol. 85:214–218, 2013. © 2012 Wiley Periodicals, Inc.