HBV-前S_2抗原和抗体在乙型肝炎中的临床意义

本文用酶免疫法检测了HBV-前S_2抗原和抗体在乙型肝炎患者血清中的变动。结果表明,在急性HBV感染时,病程一月内前S_2抗原的阳性效为86.9％,抗前S_2抗体的阳性率为27.3％;在病程1～6个月间,前S_2抗原的阳性率降至34.7％,抗前S_2抗体的阳性率升至60％。前S_2抗原在HBeAg和HBV-DNA阳性血清中的检出率(84.2％)明显高于在HBeAg和HBV-DNA阴性血清中的检出率(20.0％),而抗前S_2抗体则相反。在慢性活动性肝炎时,前S_2抗原的检出率为80％,在慢性迁延性肝炎时的检出率为82.6％,抗前S_2抗体在慢性乙型肝炎病人血清中的检出率均很低。本文结果提示前S_2抗原的表达与HBV病毒的复制相关,而抗前S_2抗体的出现可作为预示HBV感染恢复的指标。     

HBV-前S_2抗原单克隆抗体制备、特性和应用

用HBsAg制备了一株分泌乙型肝炎病毒前S_2抗原单克隆抗体(MAb)的杂交瘤细胞株(1E_2),小鼠腹水中MAb效价达10~7。1E_2MAb有很强的抑制多聚人血清白蛋白(pHSA)和HBsAg中前S_2抗原结合的能力;与重组痘菌病毒表达的中分子蛋白(含S和前S_2抗原)而不与小分子蛋白(含S抗原)反应;与人工合成的前S_2肽段(N端12O-146氨基酸序列)特异性结合。应用1E_2MAb的酶免疫法检测乙肝病毒感染者血清中前S_2抗原,表明前S_2抗原滴度和HBsAg滴度有较好的相关性,在EBeAg阳性血清中更为明显;HBeAg阳性血清前S_2抗原的阳性率和几何平均滴度明显高于HBeAg阴性/HBsAg阳性血清。     

乙型肝炎患者血清中HBV的前S_1和前S_2蛋白的检测及其意义

乙肝病毒的外壳是由S、前S_1、前S_2蛋白所组成。我们用抗前S_1和抗前S_2单克隆抗体检测106例病毒性肝炎的前S_1蛋白和前S_2蛋白,结果表明急性乙肝早期的前S_1、前S_2蛋白(n=16,75％和87,5％)明显高于急性乙肝恢复期(n=33,21.2％、12.1％);慢活肝的前S_2蛋白(n=15,93.7％)明显高于慢迁肝(n=18,55.6％),前S_2蛋白与HBsAg、eAg和HBV-DNA呈正相关(P<0.05,<0.05<0.001),支持前S_2蛋白的出现和HBV复制有关。     

HBV-DNA阳性育龄妇女HBV血清标志物、前S1抗原的检测分析

目的：探讨HBV—DNA阳性育龄妇女病毒复制与HBV标志物及前S1抗原的关系。方法：对1643例慢性乙型肝炎育龄妇女采用荧光定量PCR法检测血清HBV—DNA,酶联免疫吸附法（ELISA）检测HBV标志物和前S1抗原。结果：HBV—DNA阳性育龄妇女432例。其中,HBsAg阴性者占19．44％,前S1抗原总阳性率55．09％,且随着HBV—DNA载量增加,前S1抗原阳性率也升高。结论：采用HBV—DNA、HBV标志物、前S1抗原联检,才能更准确提供育龄妇女HBV感染诊疗依据,有效控制HBV感染率。     

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.     

乙型肝炎病毒前S蛋白前S_1前S_2与HBV DNA HBCAg的关系

采用LAB-ELISA.斑点-ELISA和斑点杂交技术检测60份急性、慢性肝病患者血清前S_1、前S_2、HBcAg和HBV DNA等标志.前S_1、前S_2和HBcAg的检出率分别为48.3%、45%和43.3%,三者无显著性差别.HBcAg阳性组前S_1,前S_2的检出率显著高于HBcAg阴性组.提示前S_1.前S_2与HBcAg关系密切;研究还表明.前S_1.前S_2与HBV DNA、HBeAg均有一定的关系.     

rIFNα-2b治疗慢性乙型肝炎中血清前S_1、S_2的动态变化

本文对5例肝活检证实的慢性乙型肝炎病人在短程强的松龙撤除后应用重组干扰素α-2b治疗过程中对血清前S_1和前S_2蛋白进行了动态观察。疗程结束时,3例病人的血清前S_1、前S_2蛋白滴度逐渐下降,尤以前S_2蛋白的下降为明显,前S_1和前S_2蛋白的下降与DNAp、HBeAg的消失相关。另2例对此治疗无反应的病人,在治疗期间,血清前S_1和前S_2蛋白无明显变化,在治疗结束时,血清HBV-DNA,DNAp及HBeAg仍阳性。结果提示慢乙肝抗病毒治疗过程中,前S_1和前S_2蛋白的测定有助于监测疗效。     

检测PHSAR的固相血凝法

<正> 现知乙型肝炎病毒表面抗原上有聚合人血清白蛋白受体(PHSAR),它由HBVS基因的前S_2区编码,是病毒复制和具有传染性的标志。常用测定方法为间接血凝、放免     

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。     

乙肝五项、HBV-DNA定量及乙肝前S1抗原联合检测用于诊断乙肝的临床价值分析

目的分析乙肝五项与病毒(HBV)前S1抗原及病毒DNA的相关性,并探讨联合检测对乙肝诊断的临床价值。方法选取本院门诊部和住院部2013年10月至2015年10月期间收治的患者作为研究对象,收集其HBsAg阳性血清,采用酶联免疫吸附法(ELISA)检测其前S1抗原和乙肝五项,采用荧光定量PCR法检测其HBV-DNA水平,对比分析不同模式下前S1抗原的表达水平和HBV-DNA的水平,并分析其相关性。结果乙肝前S1抗原阳性率与HBV-DNA阳性率在不同HBV-M模式下比较,差异无统计学意义(P>0.05)。此外1+3+5+组中HBV-DNA阳性拷贝数较高,多数高于105copies/m L;而1+4+5+、1+2+、1+3+、1+4+和1+5+组的前S1+与前S1-组相比,其HBV-DNA阳性率差异无统计学意义(P>0.05)。HBsAg阳性血清标本中前S1抗原在HBe Ag+组中阳性率明显高于HBe Ag-组,差异具有统计学意义(P<0.05),HBsAg阳性血清标本中前S1抗原与HBe Ag具有一定的相关性。HBsAg阳性血清标本中前S1抗原在HBV-DNA+组中阳性率明显高于在HBV-DNA-组,差异具有统计学意义(P<0.05),HBsAg阳性血清标本中前S1抗原与HBV-DNA具有一定的相关性。结论前S1抗原可较好反映乙肝病毒存在和复制的情况,将前S1与乙肝五项和HBV-DNA进行联合检测可较早发现较低水平病毒的感染,其可作为抗病毒疗效的评判指标,对乙肝的临床诊断和疗效评价均具有较好的应用价值。     

Pre-S1 and Pre-S2 gene-encoded proteins in liver and serum in chronic hepatitis delta infection

Frozen cryostat sections and sera from 30 patients with chronic delta infection were examined for pre-S1 and pre-S2 gene-encoded proteins, and the results were compared to markers in liver and serum HBV and HDV replication. Pre-S1 and pre-S2 were detected by indirect immunofluorescence (IF) in the liver in all 26 patients with histochemically demonstrable HBsAg. Pre-S peptides were found by double IF to have a predominantly cytoplasmic expression and to be located in the same hepatocytes expressing HBsAg. Liver cells expressing hepatitis delta antigen (HDAg) were frequently negative or very weakly positive for HBsAg and pre-S peptides, but occasional HDAg positive hepatocytes were also strongly positive for HBsAg and for pre-S peptides, particularly pre-S2. Circulating pre-S1 was detected in 24 patients (80%) and pre-S2 in 27 (90%). Detection of pre-S peptides in liver and serum was independent of HBV and HDV replication and of the HBV-DNA integration state. There was no correlation between the amount of circulating pre-S peptides and serum HBV-DNA and HDV-RNA. These results indicate that in chronic HDV infection, formation and secretion of pre-S peptides and of HBsAg occur independently of HBV and HDV replication and secretion. They further indicate that in the acquisition by replicating HDV of an HBV-derived envelope in the liver, both HBsAg and pre-S peptides are concomitantly available but circulating HDV-RNA is not invariably associated with the presence of these peptides in serum.     

Hepatitis B pre-S1 and pre-S2 proteins: clinical significance and relation to hepatitis B virus DNA

Pre-S proteins may have an important role in virus assembly and virus entry into the host cell. The presence of pre-S proteins in serum has also been thought to correlate with active viral replication. To investigate whether pre-S proteins in serum might have additional diagnostic and/or predictive value for liver sequelae in HBV infection, sera from six different serological groups of patients with HBV markers (total number 363) and different manifestations of liver histology were examined for the presence of pre-S1 and pre-S2 proteins using micro-ELISAs. Pre-S1 and pre-S2 proteins were detected significantly more often in HBV-DNA-positive than in HBV-DNA-negative sera from HBsAg carriers. However, pre-S1 and pre-S2 proteins were also found in HBV-DNA-negative HBsAg carriers irrespective of serum HBeAg/anti-HBe or liver histologic findings. These results suggest that the presence of the pre-S1 and or pre-S2 proteins in serum either does not seem to reflect the presence of active viral replication and active liver disease or pre-S proteins are more readily detectable than HBeAg and HB-DNA as measured by a dot-blot technique. Furthermore, the presence of pre-S proteins in serum is strongly correlated with that of HBsAg.     

Occurrence of pre-S1 antigen in viremic and nonviremic carriers of hepatitis B surface antigen

The proteins of viral envelope, encoded by the pre-S1 region of HBV-DNA, were measured quantitatively with enzyme immunoassay using monoclonal antibodies directed to pre-S1 epitope and correlated with the expression of pre-S2 region encoded epitope and other HBV markers. In acute HBV infection, both pre-S encoded proteins were detected in sera along with markers of viral replication and disappeared shortly before complete virus clearance while high HBsAg titers were still present. Pre-S1 antigen was present in most (95.5%) symptomatic and asymptomatic chronic HBsAg carriers. There was no correlation between the presence of pre-S1 and HBeAg or HBV-DNA in serum: 73% of sera with pre-S1 determinants were anti-HBe positive, and only 25.4% were positive for HBV-DNA. Most pre-S1 activity in sera of viremic carriers was detected in fractions of sucrose gradient containing subviral 22-nm particles, and much less in those containing infectious virions. In asymptomatic, nonviremic HBsAg carriers, pre-S1 was located only on subviral 22-nm forms. Pre-S1 positive particles had no accessible pre-S2 epitope, which is recognized specifically by monoclonal anti-pre-S2 (F124) antibody. These results show that the synthesis of the large protein of HBV envelope may occur also in the absence of active viral replication, and in these cases pre-S1 encoded sequences are on subviral particles of HBsAg. Therefore, pre-S1 is not a serologic marker of infectious virus. Disappearance of pre-S1 epitopes on HBsAg occurs only before complete clearance of the virus, and this may have potential prognostic relevance.     

New assays for quantitative determination of viral markers in management of chronic hepatitis B virus infection.

We performed a quantitative study of serum hepatitis B virus (HBV) markers, including new parameters such as pre-S1 antigen (Ag), pre-S2 Ag, and anti-HBx, in 88 chronic hepatitis B surface antigen (HBsAg) carriers. New IMx assays for HBsAg and immunoglobulin M (IgM) anti-HBc detection were also used. The population studied was composed of 65 chronic hepatitis cases (40 positive for hepatitis B antigen [HBeAg] and 25 positive for anti-HBe) and 23 anti-HBe-positive, asymptomatic HBsAg carriers. Serum HBsAg levels detected by IMx were higher in HBeAg-positive than in anti-HBe-positive HBsAg carriers (all patient subgroups included) and correlated with the serum HBV DNA level (P = 0.0001). Both pre-S1 and pre-S2 Ags were detected by enzyme immunoassays in almost all HBsAg carriers. Both pre-S1 and pre-S2 Ag titers correlated positively with the serum HBsAg concentration (P = 0.0001), but only the pre-S1 Ag titer correlated with the level of serum HBV DNA (P = 0.02). The detection of low levels of IgM anti-hepatitis B core (anti-HBc) antibodies by IMx was associated with the presence of liver disease (P = 0.05) but not with the level of viral replication. The prevalence of anti-HBx antibodies detected by the enzyme immunoassay was slightly, although not significantly, higher in patients with high levels of HBV DNA (greater than 100 pg/ml) than in patients without detectable HBV DNA (P = 0.16). In anti-HBe-positive chronic HBsAg carriers, the quantitative detection of serum HBV DNA, pre-S Ag titers, and IgM anti HBc allowed us to predict which patients suffered from chronic liver disease and/or supported viral replication (P < 0.05). In a follow-up study of eight patients undergoing antiviral therapy, the clearance of both pre-S1 Ag and HBV DNA was associated with a subsequent clearance of HBV. Therefore, the quantitative determination of HBV DNA, pre-S Ags, IgM anti-HBc may prove useful for the decision to use and the monitoring of antiviral therapy, especially in anti-HBe-positive HBsAg carriers.     

Differential effect of chronic hepatitis D virus infection on intrahepatic expression of hepatitis B viral antigen.

AIMS: To determine how chronic hepatitis D virus (HDV) infection affects intrahepatic hepatitis B virus (HBV) antigen expression. METHODS: Ninety eight liver biopsy specimens from 68 patients seropositive for total antibody to HDV were studied by immunohistochemistry, and the amount of HBV antigens was also quantified by radioimmunoassay in 12 patients and compared with 30 patients with chronic HBV infection. RESULTS: Forty nine of the 68 patients were positive for intrahepatic HDV antigen and only five were positive for HBV core antigen (HBcAg). HBV surface antigen (HBsAg) was present in 55 (80.9%) patients and was always cytoplasmic in distribution. Hepatic pre-S1 and pre-S2 expressions paralleled that of HBsAg, and were detected in 53 (77.9%) and 54 (79.4%) patients, respectively. There was no relation between the intrahepatic expression of HDV antigen and HBsAg/pre-S1/pre-S2. Follow up biopsy specimens in 25 patients showed either static or deteriorating histology while intrahepatic HDV antigen remained the same or fell. The patients with intrahepatic expression of HBcAg had either absent or noticeably decreased expression of HBcAg in their follow up biopsy specimens (median two years). In contrast, HBsAg/pre-S1/pre-S2 were the same or increased (p less than 0.001). Quantification of intrahepatic HBsAg in patients with chronic HDV infection (0.61 pg/hepatocyte, range: 0.05-1.08, n = 12) showed no difference with patients with chronic HBV infection alone (0.64 pg/hepatocyte, range: 0.02-1.02, n = 30, p = NS). CONCLUSION: These data indicate that chronic HDV infection suppresses intrahepatic expression of HBcAg but not HbsAg and pre-S antigens, suggesting a differential effect of chronic HDV infection on HBV gene expression.     

一种简易检测HBV聚合人血清白蛋白受体P-R-P酶法的建立与临床应用

本文介绍了一种简易检测这种受体的P-R-P酶法及其临床应用,它是用针对受体的酶标物,即以改良的过碘酸钠法,用辣根过氧化物酶标记的聚合人血清白蛋白(pHSA)作为标记物,采用夹心法原理来检测患者血清中受体的,可排除原有方法中一些非特异性干扰。通过对286份临床标本的检测及与现有酶联法(以抗HBs作酶标物)比较,具有微量、快速、敏感及特异等优点。两种酶法的相对符合率,灵敏度和特异性,分别为95.6％,92.1％和98.5％,并对结果和两种酶法在检出率上的差异作了讨论。此法在肝炎诊断,鉴别诊断以及乙肝发病机制研究上具有较高的实用价值。     

慢乙肝患者HBV-DNA和Pre-S1Ag及HBVM与肝脏功能的检测意义

目的:了解乙型肝炎病毒HBV-DNA、Pre-S1Ag、乙肝标志物(HBVM)和肝脏功能之间的关系及临床意义.方法:采用荧光定量聚合酶链式反应(FQ-PCR)和ELISA分别检测169例乙肝病人血清HBV-DNA含量和乙肝标志物及Pre-S1Ag与肝功能,并对结果进行对比分析.结果:各种不同类型乙肝HBsAg的阳性率均高于91.1%,HBeAg、Pre-S1Ag的阳性率随HBV-DNA拷贝数的升高而升高,但肝功能和HBV-DNA拷贝数之间不存在相关关系.结论:同时检测血清乙肝标志物、Pre-S1Ag、HBV-DNA和肝脏功能对临床HBV感染、复制及传染性的判断以及肝功能损伤程度均有重要意义.     

Levels of pre-S antigens and HBV DNA in sera from high and low viremic HBV carriers.

The biological and clinical significances of pre-S antigens and HBV replication were investigated. Some 125 sera, 28 from HBeAg and 97 from anti-HBe-positive HBsAg, carriers were studied. The aim was to verify whether pre-S antigens could be expressed in serum in complete absence of viremia. Pre-S proteins, determined by an enzyme immunoassay, were found in sera regardless of the presence of HBV DNA, as detected by spot-hybridization. The sera without detectable HBV DNA were investigated further by PCR using specific primers for the S and C regions of HBV. PCR analysis of samples revealed that 4 out of 5 HBeAg and 33 out of 41 (80.5%) anti-HBe positive sera contained HBV-amplified sequences of S and C regions. Pre-S antigen values correlated well with the amounts of HBV DNA in serum detected by PCR in anti-HBe-positive subjects with high titers of pre-S antigens (10(4)-10(6)). In addition, PCR highlighted the presence of HBV DNA sequences in 8 out of 17 (47.1%) pre-S-negative HBsAg-positive sera.     

Overlapping gene mutations of hepatitis B virus in a chronic hepatitis B patient with hepatitis B surface antigen loss during lamivudine therapy

Disappearance of hepatitis B surface antigens (HBsAg) in chronic hepatitis B usually indicates clearance of hepatitis B virus (HBV) infection. However, false HBsAg negativity with mutations in pre-S2 and 'a' determinant has been reported. It is also known that YMDD mutations decrease the production of HBV and escape detection of serum HBsAg. Here, we report overlapping gene mutations in a patient with HBsAg loss during the lamivudine therapy. After 36 months of lamivudine therapy in a 44-yrold Korean chronic hepatitis B patient, serum HBsAg turned negative while HBV DNA remained positive by a DNA probe method. Nucleotide sequence of serum HBV DNA was compared with the HBV genotype C subtype adr registered in NCBI AF 286594. Deletion of nucleotides 23 to 55 (amino acids 12 to 22) was identified in the pre-S2 region. Sequencing of the 'a' determinant revealed amino acid substitutions as I126S, T131N, M133T, and S136Y. Methionine of rtM204 in the P gene was substituted for isoleucine indicating YIDD mutation (rtM204I). We identified a HBV mutant composed of pre-S2 deletions and 'a' determinant substitutions with YMDD mutation. Our result suggests that false HBsAg negativity can be induced by combination of overlapping gene mutations during the lamivudine therapy.