一起戊型肝炎暴发的血清流行病学调查

目的 了解一起戊型肝炎暴发的血清学特点。方法 对某单位在10d内先后发病的5例急性黄疸性肝炎患者、在该单位食堂就餐的1675人(暴发人群)及未就餐的邻近单位883人(对照人群)的血清在首发病例26d后进行抗-HEV IgM和IgG检测，数据进行统计学分析。结果 5例患者抗-HEV IgM和IgG均为阳性。暴发人群抗-HEV IgM和IgG的阳性率分别为8．7％和38．4％，而对照人群仅分别为0．1％和28．6％，差别均有非常显著意义。暴发人群145例抗-HEV IgM( )中，ALT增高32例，明显高于IgM(-)及对照；而抗-HEV IgM(-)的ALT增高比例并不高于对照人群。4例患者系列血清检测见抗-HEV IgM逐渐下降，感染后4个月多数转阴，而IgG在感染后第2～3个月达高峰，随后缓慢下降。暴发人群中抗-HEV IgM( )的IgG平均水平最高，IgM(-)而IgG( )的IgG平均水平亦明显高于对照，提示暴发人群中既往感染者受到了免疫加强。暴发人群中抗-HEV IgM( )者在性别及年龄组间差异无显著意义，但其中ALT增高男性的比例显著高于女性，而与年龄无关。结论 本次急性黄疸性肝炎的暴发由戊型肝炎病毒引起，与食源有关；抗-HEV IgM和IgG不仅可用于临床病例诊断，也可用于人群调查；感染危险性与年龄及性别无关，但男性ALT增高更常见。     

我国戊型肝炎ELISA诊断试剂的质量比较

目的对戊型肝炎(戊肝)诊断试剂进行质量评价。方法选择北京万泰生物药业(万泰),北京现代高达生物技术(高达),上海科华生物工程(科华),GeneLabs Diagnostic Inc(GeneLabs),和河南华美生物工程(华美)共5家公司生产的检测戊肝病毒IgG(HEV-IgG)抗体的ELISA试剂,分别检测戊肝诊断试剂参比品、可疑戊肝临床患者和正常人群血清HEV抗体。结果对24份阳性和30份阴性参比品检测,以万泰试剂检测符合率最高,阳性和阴性符合率分别为95.83%(23/24)和96.67%(29/30),总符合率为96.30%,其次为高达试剂87.50%(21/24)和100%(30/30),总符合率为94.44%,华美试剂87.50%(21/24)和96.67%(29/30),总符合率92.59%,GeneLabs66.67%(16/24)和100%(30/30),总符合率为85.18%,科华试剂45.83%(11/24)和100%(30/30),总符合率为75.93%。对42份可疑戊肝患者血清HEV抗体检测阳性率,高达试剂为100%(42),万泰和华美试剂均为97.62%(41),GeneLabs和科华试剂均为90.48%(38)。对230份正常人群HEV抗体检测阳性率,从高到低依次为万泰试剂31.74%(73),华美试剂为23.91%(55),高达试剂为17.39%(40),GeneLabs和科华试剂分别是7.83%(18)和3.48%(8)。结论5家试剂检测急性期戊肝患者抗体阳性率均能达90%以上,具有很高的一致率,而应用于普通人群HEV感染的调查,其抗体阳性率从31.74%~3.48%不等。不同的试剂存在假阳性或和阳性漏检现象。依本室参比品和人群血清阳性率来看,万泰试剂检出率较高。     

散发性急性戊型肝炎血清抗体动态变化和肝脏超微…

为探讨散发性戊型肝为血清抗体动态变化，应用酶联免疫法（ＥＩＡ）检测了７例急性戊型肝炎抗戊型肝炎病毒（ＨＥＶ）ＩｇＧ和ＩｇＭ抗体、并对１例２进行了肝超微结构病理检测，结果表明，发病１０天至４５天内抗ＨＥＶ－ＩｇＧ和ＩｇＭ滴度最高，发病第４０天仍有肝细胞肿胀，胞浆空化和线粒体固缩等病理变化。患者轿清抗－ＨＥＶＩｇＭ滴度在４５天后逐渐下降，２个月内全中消失，抗－ＨＥＶＩｇＭ滴度在４５天后逐渐下降，２个月     

散发性戊型肝炎临床与血清学研究

在每年的 2 0 0万急性病毒性肝炎中 ,戊型肝炎 (ＨＥ)感染占 17.2 %～2 0 %。我国ＨＥ不仅有小规模或大规模爆发流行 ,散发性病例也不在少数。其临床发病特点和血清特异性抗体的产生较甲型肝炎有明显区别。我们对收住院15 1例散发单纯ＨＥ的临床表现及血清特异性ＩｇＭ、ＩｇＧ抗体的特点总结如下。1　材料和方法1.1　研究对象　 1996年 1月～ 1998年10月收住院的 15 1例临床诊断为ＨＥ的患者。男性 131例 ,女性 2 0例 ,年龄 15～ 80岁 ,平均年龄 47.37岁 ,血清学甲、乙、丙、庚型肝炎病毒抗原、抗体指标阴性 ,抗 ＨＥＶＩｇＭ或抗 ＨＥＶ…     

戊型肝炎患者血清IgG抗—HEV动态变化

应用酶联免疫吸附试验(ELISA)检测80例散发性戊型肝炎病人的系列血清表明,IgG抗-HEV于发病后第3天即可阳转,于发病后3～14天和15～30天,IgG抗-HEV累积阳转率分别为68.75％和93.75％。多数病人IgG抗-HEV持续时间较短,发病后4～6个月和1～2年分别有62.50％和95％的病人阴转。90％的病人IgG抗-HEV水平于30天内达到高峰,多数病人IgG抗-HEV水平在急性期较高,恢复期降至低水平。     

戊型肝炎病毒Ⅰ型ORF3蛋白的表达、纯化及抗原性分析

目的　表达戊型肝炎病毒 (HEV)Ⅰ型ORF3全长基因片段 ,纯化表达产物并进行抗原性分析。方法　将Ⅰ型HEVORF3基因连接到融合表达载体pThioHisB ,IPTG诱导表达 ;Westernblot分析表达产物的抗原活性 ;用经高效液相色谱纯化的表达产物作为抗原 ,制备血清抗 HEVIgG及抗 HEVIgM诊断试剂 ,用国家参考品进行考核 ;用所得抗原检测临床血清中抗 HEV抗体 ,比较其与Genelabs试剂盒检测结果的差异。结果　含有Ⅰ型pThioHisB/ORF3质粒的菌株表达了相对分子质量(Mr)为 2 6× 10 3 的融合蛋白 ,免疫印迹表明其能与戊型肝炎患者恢复期血清发生特异性反应。国家参考品考核结果显示 ,用表达产物制备的抗HEVIgG诊断试剂阳性符合率为 10 0 % (10 / 10 ) ,阴性符合率为 96 .7% (2 9/ 30 ) ,总符合率为 97.5 % (39/ 4 0 ) ;制备的抗 HEVIgM诊断试剂阳性符合率为 83.3%(10 / 12 ) ,阴性符合率为 10 0 % (2 0 / 2 0 ) ,总符合率为 93.8% (30 / 32 )。与Genelabs公司试剂盒检测结果比较 ,抗 HEVIgG和抗 HEVIgMELISA总符合率分别为 94 .5 %和 87.0 %。结论　本实验所得全长ORF3基因工程表达产物具有良好的抗原性 ,可用于制备抗 HEV诊断试剂。     

4种戊型肝炎病毒IgM抗体诊断试剂在一起暴发疫情中的性能比较

目的 比较4种戊型肝炎(戊肝)病毒(hepatitis E virus,HEV)IgM抗体诊断试剂盒在1起暴发疫情中的诊断性能。方法 应用4种试剂检测一起戊肝暴发疫情中研究对象的急性期血清标本中的IgM抗体,并检测HEV RNA;进行统计学分析。结果 MP、科华、万泰和贝尔公司试剂的HEV IgM抗体检出率分别为40.00%(108/270)、29.26%(79/270)、28.52%(77/270)和20.37%(55/270),一致率为78.52%(212/270);对HEV RNA阳性血清标本,MP、科华和万泰试剂的检测灵敏度均为66.67%(32/48),贝尔试剂灵敏度为58.33%(28/48);对急性戊肝临床诊断病例血清标本,MP、科华和万泰试剂检测灵敏度为69.81%(37/53),贝尔试剂灵敏度为60.38%(32/53);贝尔与科华、贝尔与万泰试剂对HEV RNA阳性或急性戊肝临床诊断病例血清标本的HEV IgM抗体检出率差异具有统计学意义(McNemar检验：P<0.05),而其他试剂的检出率之间差异无统计学意义(McNemar检验：P>0.05);对H...     

戊型肝炎病人和实验感染戊型肝炎病毒的猕猴血清抗——H…

应用人工合成的戊型肝炎病毒（ＨＥＶ）基因组开放读码框架２（ＯＲＦ２）和３（ＯＲＦ３）两段多肽，分别建立了酶联免疫试验（ＥＩＡ），检测８５份实验感染ＨＥＶ的猕猴和１４３份戊型肝炎（简称戊肝）病人血清，结果两组抗－ＨＥＶ ＯＲＦ２阳性率分别为７０．８９％和６８．５３％，均显著高于抗－ＨＥＶ ＯＲＦ３，且前者阳转时间较早，持续阳性时间也较长。虽然多数（９７．３％）抗－ＨＥＶ ＯＲＦ３阳性血清，抗－ＨＥＶ     

散发性急性戊型肝炎血清抗体动态变化和肝脏超微结构的病理观察

为探讨散发性戊型肝炎血清抗体动态变化，应用酶联免疫法（ＥＩＡ）检测了７例急性戊型肝炎患者抗戊型肝炎病毒（ＨＥＶ）ＩｇＧ和ＩｇＭ抗体，并对１例患者进行了肝超微结构病理检测。结果表明，发病后１０天至４５天内抗ＨＥＶ－ＩｇＧ和ＩｇＭ滴度最高，发病第４０天仍有肝细胞肿胀，胞浆空化和线粒体固缩等病理变化。患者血清抗－ＨＥＶＩｇＭ滴度在４５天后逐渐下降，２个月内全部消失，抗－ＨＥＶＩｇＧ消长情况与ＩｇＭ相似，但在７个月时仍有３例（４３％）抗体阳性。     

戊型肝炎病毒IgM抗体的检测及临床意义

采用合成多肽抗原建立了检测戊型肝炎病毒－ＩｇＭ的酶联免疫吸附试验方法，成本低，操作简便，快速，重复性好，特异性强。与抗甲型肝炎病毒，乙型肝炎病毒，丙型肝炎病毒的ＩｇＭ抗体无交叉反应，排除了ＲＦ的干扰，且可被β－巯基乙醇抑制而不起反应。     

Development of a fluorescent microbead-based immunoassay for the detection of hepatitis E virus IgG antibodies in pigs and comparison to an enzyme-linked immunoassay

Swine hepatitis E virus (HEV) is a zoonotic virus and pigs are considered as an important reservoir. Swine HEV infection is widespread and most pig herds are infected. Humans can be infected with swine HEV via consumption of undercooked pork or through direct contact with infected pigs. To minimize the risk of zoonotic transmission, sensitive tools to assess the HEV infection status of pigs and pork products are needed. The objective of this study was to develop a fluorescent microbead-based immunoassay (FMIA) for the detection of IgG antibodies against swine HEV and compare it to an in-house enzyme-linked immunoassay (ELISA). Three sets of samples were utilized: (A) samples from pigs infected experimentally with different strains of HEV (positive controls, n = 72), (B) samples from known HEV-negative pigs (negative controls, n = 62) and (C) samples from pigs of unknown HEV infection status (n = 182). All samples were tested by both ELISA and FMIA. The results on the experimental samples with known HEV exposure indicate that both assays have a specificity of 100% while the sensitivity ranges from 84.6% (ELISA) to 92.3% (FMIA). The overall prevalence of HEV IgG antibodies in field samples from pigs with unknown HEV exposure was 21.9% (40/182) for the ELISA and 21.4% (39/182) for the FMIA. The two assays had an almost perfect overall agreement (Kappa = 0.92).     

Serological diagnostics of hepatitis E virus infection

Development of accurate diagnostic assays for the detection of serological markers of hepatitis E virus (HEV) infection remains challenging. In the course of nearly 20 years after the discovery of HEV, significant progress has been made in characterizing the antigenic structure of HEV proteins, engineering highly immunoreactive diagnostic antigens, and devising efficient serological assays. However, many outstanding issues related to sensitivity and specificity of these assays in clinical and epidemiological settings remain to be resolved. Complexity of antigenic composition, viral genetic heterogeneity and varying epidemiological patterns of hepatitis E in different parts of the world present challenges to the refinement of HEV serological diagnostic assays. Development of antigens specially designed for the identification of serological markers specific to acute infection and of IgG anti-HEV specific to the convalescent phase of infection would greatly facilitate accurate identification of active, recent and past HEV infections.     

人源抗乙肝表面抗原基因工程抗体的研究

目的 研制人源抗乙肝表面抗原(HBsAg)基因工程抗体.方法 采集多个HBsAg加强免疫后表面抗体阳性(HBsAb+)志愿者的外周血,分离淋巴细胞,构建人源抗HBsAg Fab噬菌体抗体基因文库.用纯化的HBsAg对抗体库进行富集筛选,经过序列测定确定抗体轻重链型别,分别克隆入真核细胞表达载体pAc-FcR和HL51-14,转染昆虫Sf9细胞和293T细胞,利用杆状病毒/昆虫细胞系统和哺乳动物细胞系统实现全抗体的分泌型表达.结果 成功筛选出20株抗HBsAg的人源Fab抗体并制备出其中6株的IgG全抗体,竞争性ELISA结果显示6株全抗体针对的是HBsAg 3个不同表位.结论 成功筛选并获得了6株针对3个不同表位的抗HBsAg的IgG全抗体,为治疗性抗体和新疫苗的研制奠定了基础.     

Differences in capabilities of different enzyme immunoassays to detect anti-hepatitis E virus immunoglobulin G in pigs infected experimentally with hepatitis E virus genotype 3 or 4 and in pigs with unknown exposure

Hepatitis E virus (HEV), a major cause of acute viral hepatitis in humans in many developing countries, is highly prevalent in the pig population worldwide. The objective of this study was to assess the capability of three porcine prototypes of a human enzyme-linked immunosorbent assay (ELISA), an in-house ELISA and a line-immunoassay (LIA) to detect anti-HEV antibodies in pigs infected experimentally with HEV (n = 57), known to be negative for HEV infection (n = 27), or with unknown exposure to HEV infection (field samples, n = 90). All 27 samples from non-infected pigs were negative with all five assays. The earliest detection of anti-HEV antibodies occurred at 14 days post-inoculation (dpi) with four of five assays. From 42 dpi, all samples from infected pigs were detected correctly as anti-HEV positive. Kappa analysis demonstrated substantial agreement among tests (0.62-1.00) at 14 dpi and complete agreement (1.00) at 56 dpi. The overall area under the curve for all quantitative tests as determined by receiver operator characteristic analysis ranged from 0.794 to 0.831 indicating moderate accuracy. The results showed that all five assays can detect anti-HEV IgG antibodies accurately in pigs infected experimentally with HEV. In field samples, a higher prevalence of anti-HEV IgG was found in breeding herds than in growing pigs (100% versus 66.7-93.9%). These serological assays should be very useful in veterinary diagnostic labs for HEV diagnosis in swine.     

人工合成戊型肝炎病毒抗原多肽片段的特性研究

利用固相合成肽法,参照计算机对戊肝病毒抗原结构的模拟分析,自行选择合成了7个HEV肽段,分别为EH174,EH370,EH362,EH352,EH232,EH286和EH276。建立检测戊型肝炎病毒抗体(抗HEV)的酶联免疫测定法(ELISA),检测合成肽段与戊肝患者血清的反应性,结果表明:(1)多肽抗原EH362,EH370抗体检出率为86.7％,EH286和EH276的检出率分别为46.7％和26.7％;(2)多肽抗原专一性强,与正常人血清、正常小鼠血清及单纯甲型、乙型、丙型肝炎抗体阳性血清无交叉免疫反应;(3)与88份曾经Generabs药盒检测抗-HEV的血清对照比较检测,该ELISA法与Genelabs药盒的结果符合率为95.5％。对合成肽段进行分子修饰和化学处理。大大提高合成肽与戊肝患者血清的敏感度和反应率。为合成肽用于建立对戊肝感染进行特异诊断的ELISA法提供了有利的条件。     

乙型肝炎病毒e抗原的纯化及其在人血清抗体检测中的应用

目的方法利用ＺｈｏｕＪｉａｎ教授提供的重组质粒ＤＮＡｐＢＶ２２０／ｅＡｇ，转染ＤＨ５α细胞，经３０℃培养３小时，４２℃培养６小时温度诱导后，提取乙型肝炎病毒ｅ抗原（ＨＢｅＡｇ）。经ＤＥＡＥ－ＳｅｐｈａｒｏｓｅＦａｓｔＦｌｏｗ层折纯化后免疫打点分析，收集阳性蛋白。经ＳＤＳ－ＰＡＧＥ电泳，考马斯亮蓝Ｇ２５０染色，显示为单一的蛋白带，用免疫印迹法（Ｗｅｓｔｅｒｎｂｌｏｔ）和免疫打点（ｌｍｍｕｎｏｄｏｔ）法，确定表达产物的特异性。将纯化的ＨＧｅＡｇ用酶联免疫吸附试验（ＥＬＩＳＡ）和免疫条。方法基检测人血清中的相关抗体。结果经４９例乙型肝炎病毒ｅ抗体（ＨＢｅＡｂ）阳性病人血清检验证实了其特异性。结论免疫条方法较ＥＬＩＳＡ方法更敏感、特异。     

人禽流感酶联免疫诊断方法的建立及其初步应用

目的建立人禽流感H5抗体的酶联免疫检测方法（ELISA）。方法用ELISA法检测疑似高致病性禽流感患者及正常人群血清标本中H5IgG抗体。结果23例疑似高致病性禽流感患者血清样品中有4例H5IsG抗体阳性，阳性率为17．40％，与血凝抑制试验（HI）和微量中和试验（NI）相比较，三者符和率为100％。234例正常人群血清H5 IgG抗体检测均为阴性。结论建立的ELISA方法可用于高致病性人禽流感的血清样品初步筛查。     

Circulating IgE levels in patients with gynecological tumors

Summary Serum IgE levels in 148 women with a histological confirmed genital cancer of different stage and site and from 23 patients with benign ovarian tumors were determined prior to therapy. For determination of IgE an enzyme — linked immunosorbent assay was used.Result obtained did not give evidence that circulating IgE levels are different to healthy control subjects. Findings were not influenced by tumorlocalization, by primary tumor or recurrence.     

HBsAg ELISA试剂盒筛检结果的评估

目的 评估部分国产与进口HBsAg ELISA试剂盒筛查血源的价值.方法 选用部分国产和进口HBsAg ELISA试剂,对中国生物制品检定所120份HBsAg临床科研组合血清样本及随机抽取400份东莞市无偿献血者血清标本为验证样本进行同步平行检测,以中国生物制品检定所组合血清及献血者HBsAg阳性并经中和试验证实者为金标准.结果采用四格表法计算两者对等指标并进行评价.结果 国产HBsAg ELISA试剂和进口HBsAg ELISA试剂的灵敏度分别为85.71%（72/84）和100%（84/84）;特异性分别为100%（436/436）和96.55%（421/436）;尤登指数分别为0.86、0.97;两种试剂重复合格率均为100%.结论 试剂的灵敏度以进口HBsAg ELISA试剂较好,但特异性比国产试剂差,两种试剂的重复性均好,两种试剂联合筛检献血者血样标本可提高临床输血的安全性.