乙型肝炎患者血清抗HBs独特型抗体免疫复合物的检测

用聚乙二醇沉淀，胰蛋白酶处理乙型肝炎（乙肝）患者血清后，再用酶联免疫吸附法检测检样中的含抗ＨＢｓ独特型抗体免疫复合物（抗ＨＢｓ－Ａｂ２－ＩＣ）。结果发现，乙肝患者血清中ＩｇＧ，ＩｇＭ类抗ＨＢｓ－Ａｂ２－ＩＣ检出总阳性率分别为：急性乙型肝炎１４．５％（９／６２），慢性活动性肝炎２１．９％（２８／１２８），慢性迁延性肝炎１２．５％（２／１６）。抗ＨＢｓ－Ａｂ２－ＩＣ阳性者的抗ＨＢｓ独特型抗体检出率８４     

淮南矿区乙型肝炎患者循环免疫复合物检测

我国为乙型肝炎高发区,人群调查表明,仅乙型肝炎病毒标志物检测HBV-M的检出率高达58.4％,HBsAg携带率为10％,而且乙型肝炎易迁延或慢性化,进而少数病例转变为肝硬化或肝癌,对健康危害极大,乙型肝炎的临床表现呈多样化,其病毒并非仅存在肝脏内,在脾脏和血细胞中均有病毒存在,致病机制复杂,为探讨循环免疫复合物(CIC)在乙肝发病机制上的临床意义,我们在进行乙肝系列研究的同时进行了正常人群(对照组)和乙型肝炎患者(检测组)循环免疫复合物的对比检测,现分析报道如下。     

乙型肝炎患者前S2抗原,抗体检测与临床

近几年来，有关乙型肝炎病毒前S2蛋白研究取得了突破性进展。乙型肝炎病毒包膜由蛋白质、碳水化合物及脂质组成，除了HBsAg作为主要蛋白外，还有另外两种蛋白，这两种蛋白由DNA序列编码，位于S基因框架内，即前S1蛋白和前S2蛋白，它们与HBV感染与复制有着密切的关系，可作为急性HBV感染的一个新的检验系统。1检测对象与方法1．1检测对象66例血清标本均选自我院门诊及住院病人，经ELISA两对半检查，并结合临床诊断确诊的乙肝患者，另外还选入甲肝病人20例，丙肝病人10例，均符合第六次全国病毒性肝炎会议标准。血清分离后－20℃保存…     

胶固素ELISA检测流行性出血热循环免疫复合物

以牛胶固素建立固相酶联免疫吸咐试验用于检测循环免疫复合物（ＣＩＣ）。对２２８份流行性出血热、４０份慢性肝炎及３０份自身免疫性疾病患者血清标本进行检测，ＣＩＣ检出率分别为８０．２６％、８０．００％及８６．６６％，其相对平均含量分别为２１．４５±５．５６ｍｇ／Ｌ、３８．６６±７．２１ｍｇ／Ｌ及１５．９８±４．３４ｍｇ／Ｌ。胶固素用于检测ＣＩＣ不受ＤＮＡ、内毒素及肝素等影响，试验方法简便、敏感是其优点。     

乙型肝炎患者血清自身抗体检测的研究

目的　检测乙型肝炎患者血清自身抗体并分析其临床意义。方法　以 HEP- 2细胞、鼠胃和鼠肾组织为抗原 ,采用间接免疫荧光法对 1 1 7例乙型肝炎患者血清及 30例正常人血清作抗核抗体、抗平滑肌抗体和抗线粒体抗体检测。结果　乙型肝炎患者自身抗体总阳性率为 1 8.8% ,高于正常对照组 ,差异有显著性 (χ2 =4.1 9,P<0 .0 5 )。自身抗体以低滴度为主 ,多见于抗平滑肌抗体和抗核抗体。结论　乙型肝炎患者存在多种自身抗体 ,观察其自身抗体的滴度与类型对乙型肝炎患者的治疗有一定的参考价值。     

滴金免疫法检测血清抗HBcIgM及IgG抗体

目的为简便快速地进行抗ＨＢｃＩｇＧ和ＩｇＭ测定。方法，用抗人ＩｇＭ或抗人ＩｇＧ抗体包被硝酸纤维素膜，用胶体金标记抗ＨＢｃ单克隆抗体，以重组ＨＢｃＡｇ溶液作为标本中抗ＨＢｃ与金标记抗ＨＢｃ的联接剂，建立了滴金免疫法（滴金法）。结果，经与酶免疫法对比测定，二法抗ＨＢｃＩｇＭ符合率为９７．２％，抗ＨＢｃＩｇＧ符合率为９５．８％，另以相同模式在同一硝酸纤维膜上分别点加抗人ＩｇＭ和抗人ＩｇＧ抗体，同步检测抗     

乙肝患者血清中HBV—PreS2抗原和抗体的检测及临床意义

前S2（PreS2）蛋白是1981年TioUais等[1]首先发现由HBsAg编码区的前S2编码的蛋白质,它与乙肝感染和复制的关系日益受到重视,有研究表明在HBV活动性复制时肝细胞上往往有PreS2蛋白抗原表达[2],急性HBV感染恢复期首先有抗-PreS2产生,并具有保护作用[3]。本文用EIA法检测了急、慢性乙肝患者血清中PreS2抗原和抗体,并探讨与其他肝炎血清标志的关系及临床意义。1材料和方法1．l标本来源：万例急性乙型肝炎（CAH）,11例慢性迁延性肝炎（CPH）患者均选自无锡市第一人民医院和传染病医院,临床均符合lop年上海第六病毒性肝炎会讨…     

乙型肝炎病毒感染者血清抗核抗体的研究

目的探讨乙型肝炎(简称乙肝)病毒(HBV)感染者血清抗核抗体(ANA)特征及其与临床的相关性。方法收集慢性乙肝(CHB)、乙肝后肝硬化(LC)、肝癌(HCC)患者共376例,采用酶联免疫吸附试验检测ANA。结果 (1)376例患者中ANA阳性99例(26.3%),其中CHB组58例,占21.0%(58/276),LC组20例,占38.5%(20/52),HCC组21例,占43.7%(21/48);健康对照组ANA阳性2例,占3.0%(2/66)。3组均明显高于健康对照组;LC、HCC与CHB组比较,差异有统计学意义(P<0.01);LC组与HCC组比较,差异无统计学意义(P>0.05)。(2)CHB、LC和HCC组ANA以低滴度(1/100)为主,分别为72.4%(42/58)、65.0%(13/20)和71.4%(15/21)。(3)ANA滴度1/100和大于或等于1/320的丙氨酸氨基转移酶、天门冬氨酸氨基转移酶、HBV-DNA各指标比较,差异无统计学意义(P>0.05)。结论 HBV感染者出现的自身抗体以ANA为主,HCC组阳性率最高。70.7%的HBV感染者ANA为低滴度阳性,对其滴度的高低与肝细胞损伤及HBV-DNA复制率无明显相关。     

HBeAg/Anti-HBe circulating immune complexes in patients chronically infected with hepatitis B virus

Summary An enzyme-linked immunosorbent assay based on the ability of polyethylene-glycol (PEG) fixed on a solid support to adsorb circulating macromolecules (PEG-solid phase test) was developed in order to provide evidence for the existence of immune complexes of HBeAg/anti-HBe (HBeAg/anti-HBe complex) in sera of HBsAg chronic carriers. The method can detect HBeAg in immune complexes whether antigen or antibody is in excess. In the chronic phase of HBV infection, HBeAg/anti-HBe complexes are formed transiently in the course of the disease, unrelated to the phases of virus replication or peaks of hepatocytolysis, or to the histologic picture of liver disease. Our study indicates that this method offers a new approach to the understanding of biological and clinical problems of the HBeAg/anti-HBe antigenic system in chronic HBV infection.     

慢性乙肝血清标志物与HBV-DNA水平的关系

目的探讨乙型肝炎病毒免疫标志物不同模式与血清HBV-DNA定量的关系。方法用荧光定量聚合酶链式反应检测技术对619例慢性乙肝病毒感染患者血清进行HBV-DNA定量测定。结果HBsAg( )/HBeAg /抗-HBc( )组、HBsAg( )/抗-HBe( )/抗-HBc( )组、HB-sAg( )/抗-HBc( )组、单项抗-HBc( )组及抗-HBs( )组HBV-DNA阳性率分别为94.44%、32.26%、24.71%、7.14%和2.50%,HBsAg( )/抗-HBs( )/抗-HBe( )/抗-HBc( )组阳性率为15.79%。各组之间差异有统计学意义(P<0.01)。PreS1Ag 、PreS2Ag 在HBsAg( )/HBeAg(-)、HBsAg( )/HBeAg 的患者血清HBV-DNA阳性率分别为41.85%、91.64%和18.36%、46.43%。结论HBeAg、PreS1Ag与HBV-DNA高度相关,可作为HBV复制的免疫指标,HBsAg( )/抗-HBs( )同时存在可能与HBV基因变异或不同亚型二次感染有关,HBeAg 的S/CO值与HBV-DNA定量有相关性,HBsAg( )的S/CO值与HBV-DNA定量无显著相关。     

应用胸腺肽和大剂量疫苗治疗成人乙肝疫苗免疫失败的观察

目的探讨胸腺肽和大剂量疫苗提高接种乙肝疫苗免疫应答的效果。方法收集成人乙肝疫苗免疫失败的抗-HBs阴性者64例,随机分为A组和B组。A组(31例)为胸腺肽试验组,即使用胸腺肽联合接种大剂量乙肝疫苗,B组(33例)为大剂量乙肝疫苗单纯接种组。结果 A组有23例抗-HBs阳性,阳转率为82.14%(23/28),B组有17例抗-HBs阳性,阳转率为54.84%(17/31),两组阳转率差异有显著性(P<0.05)。结论使用胸腺肽以及大剂量乙肝疫苗可有效诱导乙肝疫苗免疫无应答者产生特异性免疫反应,能显著提高成人乙肝疫苗免疫应答。     

浙江地区婴幼儿和学龄前儿童乙型肝炎病毒血清学标志物的检测与分布特征研究

目的 调查和分析浙江地区婴幼儿和学龄前儿童乙型肝炎(简称乙肝)病毒血清学模式及其与年龄、性别相关的分布特征,为当地儿童乙肝的预防与控制提供依据.方法 采用电化学发光技术在Roche e601免疫分析仪上检测乙肝病毒血清学标志物.应用实验室信息管理系统(LIS)将2006年7月至2010年3月期间所有乙肝病毒血清学检测结果导出为EXCEL数据文件,通过排除重复检测者、年龄＞7周岁和住址非浙江地区者,最终筛选出24 892名研究对象,男15 810名,女9082名.采用SPSS 13.0和EXCEL 2003对乙肝病毒血清学检测结果进行统计分析.结果 乙肝疫苗平均覆盖率在浙江地区15 413名男孩和8 840名女孩中分别为99.16%(15 413/15 543)和99.17%(8 840/8 914).所有24 892名7岁内儿童中检出乙肝病毒血清学模式17种,最主要模式为"单antiHBs(+)",占65.997%(16 179/24 892);其次为"阴性"、"anti-HBc(+)anti-HBs(+)"和"anti-HBe (+)anti-HBc(+)anti-HBs(+)",依次占18.231%(4 538/24 892)、9.911%(2 467/24 892)和4.395%(1 094/24 892);其他13种低频模式检出率为0.004%～0.992%."单anti-HBc(+)"和"anti-HBe(+)、anti-HBs(+)"两种模式在男女间的检出率差异有统计学意义(χ2值分别为7.143、8.123,P均＜0.01＝,其余15种模式性别间差异无统计学意义(P均＞0.05).共检出HBsAg阳性者94名,占0.38%.总anti-HBs阳性儿童19 899名,占79.94%,其中"单anti-HBs(+)"16 178名,占总anti-HBs阳性者的81.30%.单anti-HBs阳性率最高为＞1～2岁组(87.27%),最低为2 d～2周组(13.13%);16 178名儿童在3种抗体浓度组(10～100、＞100～1 000和＞1 000 mIU/ml)的比率依次为23.64%、27.66%和13.70%.总的anti-HBc(+)儿童有3 904名,占15.68%,阳性率最高为2～4周组(57.63%),最低为3～4岁组(3.46%).1 565名"阴性"模式儿童中,阴性率最低为9～12个月组(3.25%),最高为6～7岁组(36.21%).结论 本地区学龄前儿童中"单anti-HBs(+)"检出率最高,其抗体浓度多位于100～1 000 mIU/ml间;其次为"阴性"模式.HBsAg的检出率很低.乙肝模式检出率在本地区儿童男女间无差异,而与年龄相关.分析1岁内(主要为3个月内)儿童乙肝血清学模式应考虑母亲血清乙肝抗原抗体的影响.

Abstract：

Objective To investigate the serological patterns of hepatitis B, understand the prevalence of hepatitis B infection in neonates, infants and preschool children based on hospital data in Zhejiang province and provide a basis for prevention and control of local pediatric hepatitis B infection. Methods Five serological markers were detected on electrochemiluminescence analyzer Roche E601. The results of all serological markers in children with hepatitis B from 2006 to 2010 were exported from laboratory information system and 24 892 (15 810 boys and 9 082 girls) cases were selected for further analysis according to the exclusion criterion including duplicate analysis, children older than 7 years and children living out of Zhejiang province. SPSS13.0 software and EXCEL 2003 were employed for statistical analysis in this study. Results Average HBV vaccination coverage rates among 15 413 boys and 8 840 girls were 99. 16%(98. 87% -99. 45%) and 99. 17% (98. 29% -99. 53%), respectively. Seventeen serological patterns of hepatitis B were observed, and "anti-HBs(+)alone"(64. 997%) was the dominant patterns followed by "negative" (18. 231%), "anti-HBc(+)anti-HBs(+)"(9.911%) and "anti-HBe(+)antiHBc(+) anti-HBs(+)"(4.395%), and the positive rates for other patters were 0. 004% -0. 992%. There were significant difference of detection rates for "anti-HBc(+)alone" and "anti-HBe(+)anti-HBs(+)"patterns between male and female (χ2=7.143,8.123, all P＜0.01), and there were no significant difference of detection rates for the other 15 patterns between male and female (all P ＞0.05). Ninety-four HBsAg positive cases were detected, accounting for 0.38%. Totally 19 899 anti-HBs positive cases were detected, accounting for 79.94%. Totally 16 187 cases were "anti-HBs(+) alone", accounting for 81.3%of the total anti-HBs(+) cases. The group of 1-2 years and 2 day-2 week had the highest and lowest rate for anti-HBs(+)alone respectively. The ratios in three groups with different anti-HBs levels (10-100 mIU/ml,100-1000 mIU/ml and ＞ 1000 mIU/ml) in 16 178 anti-HBs(+) children were 23. 64%, 27.66% and 13. 70%, respectively. Totally 3 904 cases with anti-HBc(+) were detected, accounting for 15.68%. The highest positivity rate and lowest positivity rate were in the group of 2-4 week and 3-4 week respectively. In total 1 565 cases with negative results, the lowest negativity rate and highest negativity rate were in the group of 9-12 month (3. 25%) and 6-7 age (36. 21%) respectively. Conclusions The dominant serological patterns of hepatitis B in local young children is "anti-HBs (+) alone" and most of the anti-HBs concentrations ranged from 100 to 1000 mIU/ml, followed by "negative". The total positive rate of HBsAg is mild. The rate of hepatitis B serological patterns in children is associated with ages but independent of gender. For interpretation of serological patterns of HBV in children below 12 months (especially in 3 months) the status of their mothers' HBV infection should also be taken into consideration.     

初诊慢肝患者血清中乙肝病毒DNA载量与血清标志物的关系

目的探讨初诊慢性乙型肝炎血清标志物与HBV DNA的关系。对象与方法采用酶联免疫方法和荧光定量聚合酶链反应分别对512例不同乙肝病人及202例HBeAg阴性、257例HBeAg阳性慢肝病人进行血清标志物、HBV DNA定量检测并分析结果。结果慢肝组有315例(68.9%)病毒颗粒浓度高于107,肝硬化组7例(13.2%)病毒颗粒浓度高于107;HBeAg阳性慢肝组245例(95.3%)患者病毒颗粒含量高浓度,HBeAg阴性慢肝组68例(33.7%)患者病毒颗粒含量在105-7copy/mL区段。结果慢肝组与肝硬化组相比,高浓度病毒以慢肝组中最明显,病情发展至肝硬化期病毒含量多数不高;慢肝病人中不同乙肝血清标志物模式其病毒含量分布特点不同,与HBeAg阳性慢肝组相比HBeAg阴性组体内病毒复制水平低(p<0.01),传染性较前者弱。最常见前C区突变,导致HBeAg阴性CHB病情加重和病毒复制增加。     

HBsAg阳性孕妇产前应用乙型肝炎免疫球蛋白提高婴儿对乙型肝炎疫苗的免疫反应

目的:探讨HBsAg阳性孕妇在孕晚期应用乙型肝炎免疫球蛋白(hepatitis Bimmunog lobu-lin,HBIG)可否改善其婴儿对乙型肝炎疫苗(乙肝疫苗)的免疫应答能力。方法:291例HBsAg阳性孕妇,分为HBsAg、HBeAg均阳性的A组(83例),HBsAg阳性、HBeAg阴性的B组(208例),按患者知情同意的原则,A组59例、B组116例在孕晚期应用HBIG(200U,每4周1次),为HBIG组,其余孕晚期未应用HBIG为非HBIG组。两组孕妇所生婴儿均在出生当日、1月龄分别注射乙肝疫苗5μg。检测6月龄婴儿外周血的乙型肝炎病毒血清标志物,比较与分析各组6月龄婴儿的乙型肝炎病毒携带率及抗-HBs阳转率。结果:①非HBIG组的婴儿6月龄抗-HBs阳转率在A、B组分别为38%及64%,两组间比较差异有统计学意义(P<0·05);HBIG组6月龄婴儿抗-HBs阳转率在A、B两组分别为81%及85%,均显高于非HBIG组(P<0·05)。②孕晚期应用3次HBIG者,其6月龄婴儿HBsAg、抗-HBs阳转率与应用1~2次HBIG者比较,差异无统计学意义。结论:HBsAg阳性(尤其是HBeAg同时阳性)孕妇的婴儿对乙型肝炎疫苗的免疫反应性较低,孕晚期应用HBIG可有效提高HBsAg阳性孕妇所生婴儿对乙型肝炎疫苗的免疫应答反应。     

慢性HBV感染者低浓度HBsAg相关分子调查研究

目的 探讨低浓度HBsAg人群的分子生物学特征及流行病学意义.方法 采用PCR及基因测序的方法 对HBV慢性感染者136份低浓度HBsAg(低浓度HBsAg组)和44份高浓度HBsAg(高浓度HBsAg组)血清标本进行HBV DNA、酪氨酸-蛋氨酸-天冬氨酸-天冬氨酸(YMDD)变异检测,并分别对浓缩法HBV DNA105拷贝/L的47份低浓度HBsAg和37份高浓度HBsAg血清标本进行基因型检测,以及对直接法HBV DNA10~5拷贝/L的14份低浓度HBsAg和29份高浓度HBsAg血清标本进行S基因序列、血清型进行检测分析,S基因序列采用BioEdit软件与中国株参照序列进行比对.结果低浓度HBsAg组HBV DNA阳性率、YMDD变异率和HBV DNA对数值分别为34.6%(47/136)、0(0/136)和6.5±1.4,高浓度HBsAg组分别为84.1%(37/44)、9.1%(4/44)和8.9±1.8,两组之间差异有统计学意义(浓缩法χ~2=30.8,P<0.05;直接法χ~2=53.5,P<0.05;YMDD变异率精确概率法,P=0.003;HBV DNA对数值t=6.5,P<0.05);47例低浓度HBsAg病例中分别检出B基因型16例、C基因型5例、未分型26例,14例血清型分别为adw 7例、ayw4例、adr2例、ayr 1例,在两组人群中基因型的分布差异有统计学意义(χ~2=13.5,P<0.05),血清型的分布差异无统计学意义(χ~2=4.7,P>0.05),S基因测序结果未发现S基因变异,但6处16例次存在核苷酸碱基差异而氨基酸同义的多态性特征.结论 低浓度HBsAg人群HBV DNA存在低复制现象,基因型、血清型分别以B型、adw/ayw为主,S基因呈多态性特征,低浓度HBsAg存在可能与HBV S基因特殊的分子生物学特征使HBsAg表达低下有关,或与患者感染HBV后机体免疫系统的个体反应导致低浓度HBsAg诱导机体免疫耐受有关.     

特异性免疫牛奶对乙型肝炎的辅助治疗研究

目的:观察免疫牛奶对乙型肝炎患者的辅助治疗作用。方法:选择乙型肝炎患者86例并分为两组,分别食用免疫牛奶和普通牛奶3个月,观察和记录患者食用前后症状(疲乏、纳差、失眠、黄疸、腹胀和便溏)与化验指标(ALT、AST、总蛋白、白蛋白和HBV-DNA)的改善情况,用秩和检验进行统计学处理。结果:免疫牛奶组与普通牛奶组患者在症状和肝功能改善方面差异有显著性,免疫牛奶有改善乙型肝炎患者症状和肝功能的作用,对乙型肝炎病毒增殖有一定的抑制作用。结论:该免疫牛奶可以用于乙型肝炎患者的辅助治疗。     

慢性HBV感染者低浓度HBsAg相关分子调查研究

Objective To investigate the molecular characteristics and epidemiological signification of patients with low-level HBsAg. Methods PCR and gene sequencing were used to detect HBV DNA and Tyr-Met-Asp-Asp(YMDD) mutant in 136 serum samples with low-level HBsAg and 44 sernm samples with high-level HBsAg. Genotyping was performed in 47 cases with HBV DNA 105 copies/L by concentration method and 37 cases with high-level HBsAg. S gene sequences and serotypes were analyzed in 14 cases with HBV DNA 105 copies/L and 29 cases with high-level HBsAg. S gene sequences were compared with the consensus sequence of Chinese strain by BioEdit software. Results The HBV DNA-positive rate, YMDD mutation rate and HBV DNA load (logarithm) in low-level and high-level HBsAg group were 34.6% (47/136), 0% (0/136), 6.5±1.4 and 84.1% (37/44), 9.1% (4/44), 8.9±1.8, respectively. There was statistically significant differences between two groups (for concentration method,χ2 = 30.8, P < 0.05; for direct method, χ2 = 53.5, P < 0.05; for YMDD mutation ratio, P = 0.003, For HBV DNA (log), t = 6.5, P < 0.05). The genotypes in low-level HBsAg group included type B (16/47), type C (5/47) and non-classified ones(26/47). There were significant differences between two groups (χ2=21.8, P <0.01). The serotypss included adw (7/14), ayw (4/14), adr (2/14) and ayr (1/14). There were significant differences in genotypes (χ2 = 13.5, P < 0.05) but not in serotypes between two groups (χ2 = 4.7, P >0.05). S gene sequencing results showed no S gnne variation was detected, but there were 6 single nucleotide polymorphisms in 16 cases, which would not result in the alternation of amino acid. Conclusions Low-replication phenomenon of HBV DNA was present in patients with low-level HBsAg. The major genotyps and serotype was type B and adw/ayw, respectively. Polymorphic variants have been found in the S gene. The existence of low-level HBsAg might be related with its own molecular characteristics resulting in low expression of HBsAg or immune tolerance induced by low-level HBsAg after HBV infection.