103例HBsAg阴性抗-HBc阳性患者的SP RIA

血清中存在HBsAg是机体受到乙肝病毒(HBV)感染的重要标志之一。血清中检出HBsAg并不表示病毒复制的程度及传染性强弱。抗-HBc为HBV感染最早出现较晚消失且长期存在的抗体,也可做为HBV感染的标志。为了解HBsAg阴转者体内HBV复制的情况,本文对103例HBsAg阴性,抗-HBc阳性者进行了分析。材料和方法一、检测对象:系我院门诊患者,既往有乙肝接触史或感染史,无明显肝功能改变,经固相放射免疫测定(SPRIA)HBsAg阴性,抗-HBc阳性,同时检测HBcAg、HBeAg和抗-HBe。二、方法:采用固相放射免疫分析法。试剂为中国同位素公司北方免疫试剂研究所提供,具体操     

监测乙肝病毒宫内感染的前S1蛋白测定

应用前S1蛋白双抗体夹心ELISA法测定269例足月分娩的母血与新生儿脐血的前S1蛋白，并与酶免法测定乙肝血清学标志的PCR法测定HBV-DNA相对照，以探讨检测母血、脐血前S1蛋白来评估HBV宫内感染的可能性。结果是：49例母血HBV抗原阳性者中检出前＆蛋白与HBV-DNA阳性者分别有19例与18例，其新生儿脐血中检出HBV抗原、前S1蛋白与HBV-DNA阳性者分别有9例、5例与6例，脐血中三种乙肝标志物阳性检出率无明显差异（P＞005）。49例母血HBV抗原阳性者中，有12例是HBsAg（＋）、HBeAg（＋）和HBcAg（＋）的，其孕妇血中前S1蛋白与HBV-DNA阳性者各10例，其新生儿脐血中检出HBV抗原、前SI蛋白与HBV-DNA阳性者分别有6例、5例与6例，脐血中三种乙肝标志物阳性检出率呈平行关系。提示测定母血、脐血前1S蛋白与检测HBeAg及HBV-DNA一样有助于判断母体内HBV复制与HBV宫内感染。     

荧光定量PCR法检测乙型肝炎病毒DNA

目的：了解不同乙型肝炎病毒(HBV)感染患者血清HBV-DNA含量与乙肝血清免疫标志物的关系并探讨其临床意义。方法：用荧光定量聚合酶链反应(FQ-PCR)方法定量检测375例不同临床类型HBV感染者及70例正常人血清HBV-DNA含量。结果：血清HBsAg阳性组与HBeAg阳性组HBV-DNA的检出率及含量均明显高于HBsAg阴性组和HBeAg阴性组，且HBeAg即使阴性，不论HBeAb阳性组或阴性组均仍有较高的HBV-DNA检出率。不同临床类型HBV感染者中均有HBV-DNA的检出。结论：定量PCR可真实反映HBV感染、复制及病程变化，对乙型肝炎临床诊断及治疗均有一定的临床意义。     

282例HBV感染无症状表面抗原阳性者血清ALT与乙肝前S1抗原的检测

目的了解乙型肝炎病毒（HBV）感染无症状表面抗原阳性者血清ALT和乙肝前S1抗原的状况。方法用ELISA法检测血清标志物（HBsAg、抗HBs、HBeAg、抗HBe、抗HBc）和乙肝前S1抗原,用日立7060全自动生化分析仪检测血清ALT。结果 282例无症状表面抗原阳性者,乙肝前S1抗原总检出率为37.6%;ALT升高87例,占总人数的30.8%;ALT升高的87例患者中前S1抗原检出率为67.8%;ALT正常的195例患者中前S1抗原检出率为24.1%,两组比较差异有统计学意义（P〈0.01）。在282例无症状表面抗原阳性者中,HBeAg阳性组,乙肝前S1抗原检出率为73.9%,ALT异常阳性率为52.2%;HBeAg阴性组,乙肝前S1抗原检出率为31.0%,ALT异常阳性率为25.6%,两组比较差异有统计学意义（P〈0.01）。结论乙肝病毒携带者不能完全排除ALT正常情况下的病毒复制;血清中出现HBeAb并不一定表示HBV复制停止。因此,对于无症状表面抗原阳性者应进行定期复查,监测其乙肝血清标志物、ALT和前S1抗原。     

HBV感染的CD68细胞在隐匿性HBV感染产妇胎盘中的分布及与宫内感染的关系

目的探讨HBV感染的外周血单个核细胞(peripheral blood mononuclear cells,PBMC)在乙肝宫内传播中的作用及机理。方法12例血清HBV DNA(-)、PBMC HBV DNA(+)产妇分娩的新生儿血清HBV DNA(+)和/或PBMC HBV DNA(+)的胎盘作为实验组,10例乙肝标志物均为阴性产妇的胎盘作为对照。采用SP法在连续切片上检测HBsAg和HBcAg在胎盘CD68细胞及各类细胞中的表达。结果8例新生儿血清HBV DNA(-)、PBMCsHBV DNA(+)胎盘绒毛间质5例CD68细胞HBsAg阳性,6例CD68细胞HBcAg阳性;毛细血管内5例CD68细胞HBsAg阳性,8例CD68细胞HBcAg阳性;滋养层细胞和血管内皮细胞均未见HBsAg、HBcAg阳性信号;2例新生儿血清HBV DNA(+)、PBMCs HBV DNA(-)的胎盘滋养层细胞、绒毛间质、毛细血管内皮细胞均有HBsAg、HBcAg的表达,而绒毛毛细血管内CD68细胞未见表达。2例新生儿血清和PBMC HBV DNA均阳性的的胎盘滋养层细胞、绒毛间质、CD68细胞和毛细血管内CD68细胞均有HBsAg、HBcAg的表达,毛细血管内皮细胞无表达。10例乙肝标志物全阴性产妇胎盘中均无阳性信号。结论HBV感染的外周血单个核细胞可作为宫内传播的载体。     

HBV前S1抗原与HBV五项血清标志物的分析及意义

目的 探讨PreS1抗原与HBV"五项"之间的相关性及临床意义.方法 EL1SA法采用全自动酶联免疫系统分析仪检测247例HBV "五项" 血清标志物和PreS1抗原.结果在247例HBsAg阳性血清中,HBeAg阳性的阳性率为34.8%,PreS1抗原阳性率为63.6%.86例HBeAg阳性标本中PreS1抗原阳性者72例,阳性率为83.7%.149例HBeAg阴性抗HBe阳性标本中PreS1抗原阳性者82例,阳性率55.0%.其中"大三阳"标本中PreS1抗原阳性率高达85.7%显著高于"小三阳"患者血清中PreS1抗原阳性率61.5%.结论 PreS1抗原是HBV感染和复制的标志,比HBeAg反映HBV复制更敏感,是HBV存在和复制较为直接的标志,对HBV"五项"检测起重要补充作用,在临床上判断HBV复制和疾病的预后有重要的参考值.     

血清HBcAg R1A临床价值的探讨

目前我国一些大中城市相继开展乙肝核心抗原(HBcAg)的放免检测.但检测结果的规律尚不甚一致.作者曾报道1989年1～10月检测的415例慢性HBV感染的血清HBcAg检出率,并就HBcAg与其他HBV标记和某些肝功能的关系作过初步研究.为进一步探讨血清HBeAg检出的规律及与其他乙肝标志的关系,以便客观评价其临床实用价值,我们又对自1991年1月～1994年3月检测的3033例HBV感染的血清HBcAc及其他HBV标志,包括部分应用PCR法检测HBV-DNA进行综合分析,现报告如下.     

HBcAg RIA在HBsAg阳性者中的临床应用

血清HBcAg在HBV感染中占有重要地位,它能反映血中Dane颗粒的存在及肝内HBV的复制,并可与其他的HBV血清学标志起互相配合和互相补充的作用。为此我院于1991年开展了此项目并对HBsAg阳性携带者在二对半的基础上加作了HBcAg的测定,现将结果报告如下。材料和方法一、检测对象:均系我院门诊及住院病人经SPRIA测定,HBsAg阳性,肝功能均正常的391例同时还检测HBeAg、抗-HBe、HBcAg、抗-HBc。二、方法: (一)采用SPRIA试剂由山东潍访三V公司提供。 (二)方法:均按说明书操作,HBcAg第三步温浴时间稍有不同由原来的45℃水浴30分钟改为2小时。     

SPRIA法检测360对母婴乙肝八项指标结果的调查分析

通过母婴配对HBV八项血清指标检测,探讨产妇HBV血清指标在不同的感染模式下,对胎儿的传染性。用固相放射免疫的方法(SPRIA),对母亲血清及胎儿脐血中的HBsAg、抗-HBs、HBcAg、抗-HBc、HBeAg、抗-HBe、PHSA-R(聚合蛋白受体)和抗-HBc-IgM等八项进行检测。结果表明,母体出现一项指标以上阳性者170例,占47.2％(170/360),将170例阳性血清归类分析,出现15种感染模式,其中HBsAg阳性者为18例,与之对应的胎儿脐血HBsAg阳性者16例,阴性者2例,说明垂直感染率高达88.8％(16/18)。此外,有2例胎儿脐血HBsAg阳性,而母亲血清仅有抗-HBc阳性。有1例胎儿脐血HBsAg阳性,但母亲HBV八项指标全部阴性。     

HBV DNA及HBV抗原在血清HBV标志物阴性肝炎肝组织中表达的研究

目的 研究HBV DNA及HBV抗原在血清HBV标志阴性的肝炎肝组织中的表达。方法 对45例HBV血清标志阴性阴性肝炎患者，进行肝组织HBV DNA的原位杂交及免疫组织化学染色检测。结果 原位杂交表明，HBV DNA阳性者7例，（阳性率15．56％），阳性信号主要存在于肝细胞的胞核中，少数位于胞浆内；免疫组化染色表明，HBsAg及HBcAg均呈阴性。结论 血清HBV标志阴性的肝炎肝组织中可检出HBV DNA，有利于提高对HBV感染的诊断。     

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。     

Anti-HBc titers in HBeAg and anti-HBe positive asymptomatic HBsAg carriers

A study was undertaken to establish markers for HBV replication in relation to HBeAg and anti-HBe. HBsAg carriers with serum HBeAg had DNA polymerase activity in the serum and HBcAg in the liver nuclei. Anti-HBe positive and anti-HBe/HBeAg negative sera lacked these markers. For anti-HBc the following geometrical mean titers were calculated: 1: 12,000 for HBeAg positive, 1:9, 100 for anti-HBe and anti-HBc positive, and 1:2,800 for anti-HBc positive anti-HBe/HBeAg negative asymptomatic HBsAg carriers. Follow up studies revealed mostly unchanged anti-HBc titers in all three groups over an observation period of ten to twenty months. Our data argue for a prolonged HBV replication in all HBsAg carrier subgroups compared to individuals with an uncomplicated acute virus-B-hepatitis. This study gives no final answer whether HBeAg negative HBsAg carriers have a continous HBV replication.     

Comparison of the Abbott Architect i2000 assay, the Roche Modular Analytics E170 assay, and an immunoradiometric assay for serum hepatitis B virus markers

Serum hepatitis B virus (HBV) markers are the most important data for epidemiological screening and clinical diagnosis of HBV infection, especially in endemic areas. We compared the results of the Roche Modular Analytics E170 assay, the Abbott Architect i2000 assay, and an immunoradiometric assay (IRMA) for HBV surface antigen (HBsAg), anti-HBV surface antigen (anti-HBs), HBV e antigen (HBeAg), and anti-HBV e antigen (anti-HBe). A number of serum samples (264, 263, 224, and 202 for HBsAg, anti-HBs, HBeAg, and anti-HBe, respectively) were studied. For samples giving discrepant results for HBeAg between methods, real-time PCR assays were performed. The concordance rates among the three methods were high for HBsAg (100%) and HBeAg (94.6), but low for anti-HBs (91.6%) and anti-HBe (82.2%). For anti-HBs, which could be measured quantitatively by the Modular E170 and Architect i2000 procedures, discrepant results were observed at low levels of anti-HBs. For anti-HBe, the positive rate was highest with Modular E170 (60.9%) followed by the IRMA kit (54.1%) and Architect i2000 (51.0%). This study shows substantial differences between the assay results by the three methods, which should be taken into account in determinations of serum HBV markers.     

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。     

Diagnostic significance of anti-HBcIgM prevalence related to symptoms in Canadian patients acutely or chronically infected with hepatitis B virus

A total of 362 sera from 295 Canadian patients were examined for HBsAg, anti-HBs, anti-HBc, anti-HBcIgM, HBeAg, and anti-HBe using commercial immunoassays. Serial samples from 70 acutely infected patients demonstrated that anti-HBcIgM may detect 10% more positives than HBsAg within 4 months after the onset of clinical symptoms, and all except two were negative for anti-HBcIgM after the fourth month. None of 66 asymptomatic (HBeAg rate 18.2%) and two of 14 (14.3%) symptomatic (HBeAg rate 64.3%) carriers of HBsAg were positive for anti-HBcIgM (P = 0.029). Elevated marker responses were measured in two symptomatic carriers for a 20-month period. Anti-HBcIgM was not detected in either 100 asymptomatic patients positive for total anti-HBc, negative for HBsAg and negative for or possessing low levels of anti-HBs, 25 patients with liver disorders not caused by HBV, or 20 healthy milk donors. In diagnostic laboratory practice this anti-HBcIgM test may be useful in the following situations: to supplement HBsAg testing, providing a theoretical 10% increase in positives within 4 months following onset of acute viral hepatitis; to replace testing for anti-HBc and anti-HBs in symptomatic HBsAg-negative patients; to confirm whether a patient is experiencing acute or chronic HBV infection or symptoms superimposed upon asymptomatic HBsAg carriage by another cause, such as nonA-nonB viral hepatitis.     

Immunohistological study of intrahepatic expression of hepatitis B core and E antigens in chronic type B hepatitis.

AIMS: To study the intrahepatic expression of hepatitis B virus (HBV) nucleocapsid antigen; and to determine the differential distribution of hepatitis B core and E antigens in chronic hepatitis B. METHODS: Hepatocyte expression of HBV nucleocapsid antigen was studied using rabbit anti-HBc, directed against both HBcAg and HBeAg; differential distribution of HBcAg and HBeAg was studied using murine monoclonal anti-HBc and anti-HBe in 120 patients with chronic hepatitis B. RESULTS: HBV nucleocapsid antigen was detected in 14 of 16 (87.5%) HBeAg seropositive patients with chronic persistent hepatitis (CPH), and in 54 of 64 (84.4%) HBeAg seropositive patients with chronic active hepatitis (CAH). Nuclear expression of nucleocapsid antigen was more prevalent in patients with CPH than in those with CAH; this was reversed in terms of exclusive cytoplasmic expression of nucleocapsid antigen (p < 0.05). Of 45 patients with nucleocapsid antigen in the nucleus, samples from 44 (97.8%) and 17 (37.8%) stained positively with monoclonal anti-HBc and anti-HBe, respectively. Of 65 patients with cytoplasmic nucleocapsid antigen, samples from 61 (93.8%) and 57 (87.7%) stained positively with monoclonal anti-HBc and anti-HBe, respectively. CONCLUSIONS: HBV nucleocapsid antigen is more prevalent in HBeAg positive patients with CPH than in those with CAH. Cellular expression of HBcAg and HBeAg in the cytoplasm is more or less the same; in the nucleus HBcAg exceeds HBeAg expression.     

不同剂量乙肝疫苗与HBIG联合免疫阻断HBV母婴传播的效果对比

目的 探讨10 μg和20 μg乙肝疫苗与HBIG联合免疫阻断HBV母婴传播的效果.方法 124例HBsAg阳性孕妇所生的婴儿随机分为两组,即10 μg乙肝疫苗组和20 μg乙肝疫苗组.婴儿于出生6h内及30 d分别注射200 IU HBIG,同时分别于出生24 h内、1个月及6个月注射3次10 μg或20 μg重组酵母乙肝疫苗.检测婴儿出生时以及1岁时血清HBV标志物.结果 两组新生儿血清HBsAg、HBeAg及抗-HBe阳性率与滴度之间差别均无统计学意义（P＞0.05）.所有新生儿血清HBV DNA水平均小于检测下限（500 U/ml）.出生12个月时,所有124例婴儿血清HBsAg和HBeAg检测结果均为阴性;血清HBV DNA水平均在检测下限以下;10 μg和20 μg乙肝疫苗组血清抗-HBs阳性率分别为90.3％和96.8％,差异无统计学意义（P＞0.05）;抗-HBs水平分别为325.5±342.2 mIU/ml和463.7±353.3 mIU/ml,后者显著高于前者（P=0.01）.而且,20 μg乙肝疫苗组产生高应答抗-HBs（＞ 100 mIU/ml）的比例显著高于10μg乙肝疫苗组（P =0.035）.结论 20 μg乙肝疫苗联合HBIG方案阻断HBV母婴传播的效果优于10 μg乙肝疫苗联合HBIG方案.     

Correlation between HBV DNA detection by polymerase chain reaction and Pre-S1 antigenemia in symptomatic and asymptomatic hepatitis B virus infections

The presence of hepatitis B virus (HBV) genome in sera from 73 symptomatic and asymptomatic HBsAg carriers was studied by the polymerase chain reaction (PCR) with primers specific for the S and C regions. Pre-S proteins of the HBV envelope were detected in serum by a specific monoclonal antibody in a double immunoradiometric assay. Out of twenty-five symptomatic patients with chronic active hepatitis (14 with HBeAg and 11 with anti-HBe), all were positive for HBV DNA by PCR, while 14/14 HBeAg and 2/11 (18%) of the anti-HBe patients were positive by dot blot hybridization. All but one anti-HBe patient (96%) carried Pre-S1 proteins. Among the asymptomatic HBsAg carriers, HBV DNA was detected by PCR in 14/14 (100%) HBeAg positive patients and in 25/34 (73%) anti-HBe positive patients. Pre-S1 proteins were found, respectively, in 14/14 (100%) and 11/22 (50%) of the same cases tested in parallel. The 20 healthy blood donors devoid of HBV markers and with normal transaminases tested were found negative for HBV DNA using PCR. Out of 12 patients who recovered from acute hepatitis B, all were found negative by PCR analysis after a mean follow up of 1 year after seroconversion to anti-HBs. When serial samples from 2 patients (one with acute hepatitis B, the other with chronic hepatitis B) were tested for the presence of HBV DNA and of Pre-S1 proteins, both markers showed parallel development.(ABSTRACT TRUNCATED AT 250 WORDS)     

不同慢性肝病患者血清中乙型肝炎病毒DNA定量检？…

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浓度及共临床意义。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ＨＢＶ ＤＮＡ浓度。结果 ＨＢＶＤＮＡ阳性的ＣＨ患者血清ＨＢＶ ＤＮＡ浓度为４．３６ｌｏｇ１０ＨＢＶＤＮＡ拷贝／５０μｌ，ＬＣ为４．     

Interplay of Cell and Serum Immunologic Markers in Chronic Persistent or Active Hepatitis B

Immunologic markers associated with hepatitis B virus (HBV) infection (HBsAg, anti-HBs, HBeAg, anti-HBe, anti-HBc, anti-δ) were tested by radioimmunoassay of serum from chronic hepatitis patients. The corresponding liver biopsy samples were examined for the presence of HBsAg, HBcAg, and δ antigen in the cells by direct immunofluorescence and by electron microscopy. Seventy patients were selected for the presence of both circulating HBsAg and anti-HBc. Comparison of chronic persistent (CPH) and chronic active (CAH) hepatitis showed a significantly greater frequency of intracytoplasmic HBsAg in CPH, especially in the absence of intranuclear HBsAg, and a greater frequency of intranuclear δ antigen and/or circulating anti-5 in CAH. The δ/anti-δ system was almost systematically associated with serum anti-HBe. At variance with HBeAg/anti-HBe, δ/anti-δ was found significantly more frequently in patients originating from Southern rather than from Northern or Central Italy. The prevalence of both these immunologic systems was related to the age of the patients.