HBV-DNA与乙型肝炎血清标志物的相关性分析

目的：探讨乙型肝炎病毒（HBV）DNA载量与其血清标志物的相关性。方法：运用荧光定量聚合酶链反应（FQ-PCR）、酶联免疫吸附实验（ELISA）分别检测503例患者HBV．DNA载量和HBV血清标志物。根据HBV血清标志物结果分为大三阳组、小三阳组、少见模式组、抗体阳性及全阴组,比较各组间HBV—DNA的阳性率及定量值。结果：在大三阳组、小三阳组、少见模式组、抗体阳性及全阴组HBV—DNA的阳性率分别为90％、65．1％、65．2％、2．0％,HBV—DNA的定量结果（10gHBV—DNA）别为6．32±1．96、2．01±1．68、3．48＋2．52f抗体阳性及全阴组阳性例数过低,不纳入统计）。大三阳组HBV—DNA的阳性率显著高于小三阳组（P〈0．05）。大三阳组、小三阳组HBV—DNA的阳性率与少见模式组比较,差异均无统计学意义（P〉0．05）,但大三阳组、小三阳组、少见模式组HBV．DNA的阳性率均显著高于抗体阳性及全阴组（P〈0．01）。HBsAg、HBeAg阳性组HBV—DNA的阳性率分别显著高于HB—sAg、HBeAg阴性组（P〈0．01）。小三阳组、少见模式组HBV．DNA载量均显著低于大三阳组（P〈0．01）,少见模式组HBV—DNA载量显著高于小三阳组（P〈0．05）。结论：HBV—DNA的阳性率与HBeAg、HBsAg相关;HBV—DNA栽量与HBV血清标志物模式相关。     

上海地区人类免疫缺陷病毒/艾滋病合并乙型、丙型肝炎病毒感染的临床流行病学研究

目的 研究人类免疫缺陷病毒1型(HIV-1)感染者中乙型肝炎病毒(HBV)和(或)丙型肝炎病毒(HCV)感染的流行现状及其特点。方法 对上海市(复旦大学附属)公共卫生中心就诊的170例HIV-1感染者采集血标本,使用微粒子酶联免疫法(MEIA)检测HIV-1感染者血清的乙型肝炎二对半(HBsAg、抗-HBs、HBeAg、抗-HBe、抗-HBc),应用酶联免疫吸附试验(ELISA)检测HCV抗体。结果 170名HIV-1感染者中,男性146人,女性24人,最小年龄16岁,最大年龄74岁,平均年龄41±13岁;HBsAg阳性患者19人(11．2％),抗-HBs阳性者87人(51．2％),HBeAg阳性者7人(4．1％)．抗-HBe阳性者49人(28．8％),抗-HBc阳性者111人(65．2％);抗-HCV阳性者77人(45．3％)。HIV-1、HBV和HCV三重感染者9人,约占5．3％。结论 HIV-1感染者HBsAg阳性率与普通人群相近,但HIV-1与HCV混合感染的比例显著高于普通人群。     

乙型肝炎病毒表面抗原在骨髓细胞中的存在及其与乙型肝炎病毒血清复制指标的关联

选择16例血清HBsAg阳性患者为实验组,19例血清HBsAg阴性患者为对照组,每一患者同时采取血清和骨髓涂片,用免疫细胞化学方法(PAP)检测骨髓涂片细胞中的HBsAg。结果,实验组3例骨髓细胞HBsAg阳性者,其血清中HBsAg滴度都很高,而且HBeAg均呈阳性。而在抗HBe阳性或HBeAg/HBeAb阴性者中均无骨髓细胞HBsAg阳性者。在5例血清HBV-DNA多聚酶阳性者中,骨髓细胞中HBsAg阳性者2例;6例多聚酶阴性者中,骨髓细胞中HBsAg阳性者仅1例。 本研究结果证明,HBV可在肝外组织细胞中测出,骨髓细胞HBsAg阳性的出现有集中于HBV高水平复制感染者中的倾向,同时更常见于HBV感染的较早时期。     

HBV感染中抗-(抗-HBs)独特型抗体的研究

1.75％聚乙二醇(MW6000)沉淀血清免疫复合物后,采用ELISA法检测上清中抗独特型抗体〔抗-(抗-HBs)〕。56例急性HBV感染患者血清IgM抗-(抗-HBs),第一周检出率最高(60.00％),1个月后大部分阴性,IgG抗-(抗-HBs)维持时间略长。IgM抗-(抗-HBs)较HBsAg/IgM和IgM抗-HBc消失早,将有利于急性HBV感染的诊断。27例CAH和29例CPH,IgM和IgG抗-(抗-HBs)阳性率约为38～52％,两型间无显著差异(p>0.5)。3例无症状HBsAg携带者和12例HBsAg血清疫苗接受者血清,抗-(抗-HBs)阴性。115例HBV感染者中,IgM抗-(抗-HBs)阳性者HBsAg阳性率(93.62)显著高于IgM抗-(抗-HBs)阴性者(52.31％),p<0.005;反之,HBsAg阳性血清IgM抗-(抗-HBs)阳性率(55.5S％)显著高于HBsAg阴性血清(9.68％),p<0.005;IgM抗-(抗-HBs)阳性和阴性血清的HBVDNA阳性率有显著差异(p<0.025),IgG反应到类似的结果。以上资料表明,抗-(抗-HBs)提示了一些HBV感染患者体内可能存在一种缺陷的反馈机制,导致抗-HBs产生不足而容许更活跃的HBV复制。本文证明了HBV感染患者血清中存在的抗-(抗-HBs)可能是共同决定簇“a”特异性的,还表明抗-HBs人抗-Id反应可能被纯化人血清HBsAg抑制,提示人抗-(抗-HBs)与抗-HBs结合的位点可能在HBsAg与抗-HBs结合位点内或在其附近,这和一些动物抗-(抗-HBs)的研究结果是一致的。     

乙型肝炎病毒前S1抗原与HBV血清标志物同时检测及临床意义

目的：了解前S1抗原与HBV血清标志物的关系。方法：采用ELSIA方法同时检测2101例乙型肝炎病毒感染者的前S1抗原和HBV血清标志物。结果：29例HBsAg( )、HBeAg( )标本中,有28例前S1抗原阳性,阳性率96．55％;784例HBsAg( )、HBeAg( )、抗HBc( )标本中,有679例前S1抗原阳性,阳性率86．6l％;276例HBsAg( )、抗HBc( )标本中,有197例前S1抗原阳性,阳性率71．38％;1012例HBsAg( )、抗Hbe( )、抗HBc( )标本中,有468例前S1抗原阳性,阳性率46．25％。结论：前S1抗原作为病毒复制的指标与HBeAg具有很好的一致性,又具有其独立的检测价值,可弥补HBV血清标志物检测的不足。     

乙肝病毒表面抗原的抗体双阳性者中病毒S区基因序列分析

为了解乙肝病毒（HBV）表面抗原和抗体双阳性患者中病毒的基因型及其HVB S区是否有变异。用放射免疫试剂检测HBsAg阳性样品中的抗－HBs抗体，用聚合酶链反应法检测双阳性样品中的HBV DNA，然后对阳性样品进行克隆和基因序列分析，并将所得序列与HBV不同基因型的代表株进行比较分析。结果显示389例HBsAg阳性样品中有10例为抗HBs抗体阳性；该10例双阳性样品中有5例为HBV DNA阳性；序列分析显示该5株HBV均为B基因型，其中4株为adw亚型，1株为adr亚型；其中有2株在S区的“a”决定簇的氨基酸发生了变异。     

抗—HBe阳性慢性乙肝临床研究

报告34例抗-HRe阳性慢性乙型肝炎(CHB),占同期住院抗-HBe阳性者的38％。这些患者于3—9年内肝炎再活动2—4次,累计80次。肝炎再活动时的临床表现及肝功损害与同期住院的HBeAg阳性CHB相似,但抗-HBe阳性CHB者肝硬化及肝癌发生比例显著高于HBeAg阳性者(P<0.05)。血清乙肝病毒标志(HBVM)检测发现,80次肝炎再活动中,38次(47.5％)有HBV活跃复制,提示HBV活跃复制是部分抗-HBe阳性CHB肝炎再活动的根本原因,另一部要考虑是其它肝炎病毒重叠感染的结果。     

乙肝病毒表面抗原和抗体双阳性者中病毒S区基因序列分析

为了解乙肝病毒(HBV)表面抗原和抗体双阳性患者中病毒的基因型及其HVB S区是否有变异.用放射免疫试剂检测HBsAg阳性样品中的抗-HBs抗体,用聚合酶链反应法检测双阳性样品中的HBV DNA,然后对阳性样品进行克隆和基因序列分析,并将所得序列与HBV不同基因型的代表株进行比较分析.结果显示389例HBsAg阳性样品中有10例为抗HBs抗体阳性;该10例双阳性样品中有5例为HBV DNA阳性;序列分析显示该5株HBV均为B基因型,其中4株为adw亚型,1株为adr亚型;其中有2株在S区的"a"决定簇的氨基酸发生了变异.     

高密市2014年人群乙型病毒性肝炎血清流行病学调查

目的了解高密市乙型病毒性肝炎(乙肝)流行现状,为完善乙肝疫苗策略提供依据。方法采用分层多阶段随机抽样方法,抽取高密市1~<5岁、5~<15岁、15~<30岁和30~<60岁常住人口497人进行乙肝血清流行病学调查,采用酶联免疫吸附试验标本进行乙肝病毒(Hepatitis B virus,HBV)血清学标志物(HBsAg、抗-HBs、抗-HBc、HBe Ag和抗-HBe)检测。结果共采集血清494份,HBV流行率为12.55%、HBsAg阳性率为1.42%、抗-HBs阳性率为52.02%、抗-HBc阳性率为12.55%,男、女性别差异无统计学意义(P>0.05);5~14岁组HBV流行率和抗-HBc阳性率均最低,且随年龄增长呈现升高趋势。各年龄组间差异均有统计学意义(P<0.01);HBsAg阳性率各年龄组间差异均无统计学意义(P>0.05);抗-HBs阳性率随年龄增长呈下降趋势,各年龄组间差异均有统计学意义(P<0.01);HBV流行率、HBsAg阳性率、抗-HBs阳性率在城乡之间差异均无统计学意义(P>0.05),但抗-HBc阳性率乡镇高于城市(P<0.05);7名HBsAg阳性患者中,15~<30岁组和30~<60岁组HBe Ag阳性各1人;15~<30岁组和30~<30岁组抗-HBe阳性分别为1人和4人。结论高密市为乙肝中度流行区,青少年和成人中HBV流行强度高于儿童,高密市实施以"新生儿Hep B接种"为主的乙肝控制策略取得了显著成效。     

Prevalence of hepatitis B viral markers in Vietnamese blood donors

Serum samples were assayed using radioimmunoassay in 573 Vietnamese blood donors living in Hano? (North Viet Nam). 66 (11.5%) subjects were HBsAg-positive. Of these 66 HBsAg carriers, 17 (25,8%) were positive for hepatitis B e antigen (HBeAg) and 43 (65.1%) for antibody to HBeAg (anti-HBe). 22 (3.8%) subjects were positive for antibody to hepatitis B core antigen (anti-HBc) alone. 402 (70.2%) subjects were positive for antibody to HBsAg (anti-HBs). This anti-HBs percentage increased with age. Only 83 (14.5%) subjects were negative for all hepatitis B viral (HBV) markers. This no HBV markers percentage decreased with age. The chi 2 test showed a non significant difference for frequencies of HBsAg, anti-HBc alone, anti-HBs but a significant one for frequencies of no HBV markers in men and women.     

In vitro production of antibody to hepatitis B core and E antigens by peripheral blood mononuclear cells in patients with chronic hepatitis B virus infection

To evaluate the relative immunogenicity of and the mechanism for production of antibody to hepatitis B core (HBc) and hepatitis B e (HBe) Ag, we investigated the in vitro anti-HBc and anti-HBe production by PBMC from 25 patients with chronic active hepatitis (CAH) (15 with HBeAg and 10 with anti-HBe) and 12 ASC (5 with HBeAg and 7 with anti-HBe) in the presence of PWM, rHBcAg, or purified HBeAg. PWM-stimulated culture produced higher titer anti-HBc (mean % inhibition +/- SD = 73 +/- 23%, p less than 0.001) than anti-HBe (34 +/- 17%). HBcAg stimulation elicited greater anti-HBc response (43 +/- 26%, p less than 0.001) than did HBeAg for anti-HBe (26 +/- 12%). Both HBcAg and HBeAg induced equivalent anti-HBe response. Anti-HBc production in response to HBcAg was higher in CAH patients (51 to 55%) than in asymptomatic carriers of hepatitis B surface Ag (22 to 28%) irrespective of their HBeAg/anti-HBe status, but reflecting serum anti-HBc value. Similar findings were noted in HBeAg-stimulated anti-HBe production for the two patient groups. In HBeAg- and anti-HBe-positive CAH, HBcAG-stimulated anti-HBc production was similar in T (1.4 x 10(6)) and B (0.6 x 10(6)) cells coculture, and B cells (2 x 10(6)) alone culture. However, in the HBeAg-stimulated culture, T plus B cells produced significantly higher titer anti-HBe than B cells alone did. These results indicate that HBcAg has a relatively higher immunogenicity in terms of antibody production as compared to HBeAg. Furthermore, HBcAg was shown to function as a T cell-dependent and -independent Ag, whereas HBeAg is T cell-dependent during chronic hepatitis virus B infection in man.     

Intradermal vaccination of adults with three low doses (2 micrograms) of recombinant hepatitis B vaccine. II. Persistence of immunity and induction of immunologic memory

Of the 110 dentists who had presented seroconversion 50 days after the intradermal application of three 2 micrograms doses of the Belgian recombinant vaccine against hepatitis B (HB), administered eight years before at an interval of one month between the 1st and 2nd doses and of five months between the 2nd and 3rd doses, 51 were included for the assessment of the persistence of immunity. None of the dentists had hepatitis or had received HB vaccine during this period. All subjects were submitted to serological tests for the detection of the following markers of hepatitis B virus (HBV) infection: HBsAg, anti-HBc, HBeAg, anti-HBe, and anti-HBs, with no HBsAg, anti-HBc, HBeAg or anti-HBe being detected. A microparticle enzyme immunoassay (MEIA) revealed the presence of anti-HBs at protective titers (> or = 10 mIU/ml) in 42 dentists (82.4%), with the anti-HBs titer being higher than 100 mIU/ml in 36 of them (70.6%) (good responders), between 10 and 100 mIU/ml in 6 (11.8%) (poor responders), and lower than 10 mIU/ml in 9 (17.6%) (non-responders). According to clinical data and serological tests, none of the dentists had presented disease or latent HBV infection during the eight years following the first vaccination. A 2 micrograms booster dose was administered intradermally to eight dentists with anti-HBs titers lower than 10 mIU/ml (non-responders) and to six dentists with titers ranging from 10 to 100 mIU/ml (poor responders); the determination of anti-HBs one month later demonstrated the occurrence of seroconversion in the eight non-responders and an increase in anti-HBs titer in the six poor responders. In summary, the present results demonstrated the prolonged persistence of protection against HBV infection and the development of immunologic memory provided by vaccination against HB--with intradermal application of three 2 micrograms doses of the Belgian recombinant vaccine at 0, 1, and 6 months--carried out eight years before in 51 dentists.     

Stability of a reference panel of lyophilized hepatitis B antigens and antibodies

A lyophilized hepatitis B working/reference panel has been prepared for use in standardization tests. This panel includes HBsAg, anti-HBs, HBeAg/anti-HBe and subtype reagents. Quantitative analysis of the HBsAg reagents indicates that at a storage temperature of -20 degrees C, only 1 log at maximum of RIA counts per minute would be lost in 95 years. After storage at -20 degrees C for 1 year, there has been no loss of reactivity in any of the tests used to detect HBsAg, anti-HBs, HBeAg/anti-HBe or subtypes.     

Quantitation of hepatitis B virus DNA in serum of patients with hepatoma

Hepatitis B virus (HBV) DNA in the serum of 31 patients with histologically confirmed primary hepatocellular carcinoma (PHC) from Malaysia and Indonesia was quantitated by densitometric scanning of autoradiograms obtained by Southern blot DNA hybridization, after electrophoresis using a 32P DNA cloned into plasmid pBR325 as a probe. This quantitation after electrophoresis is more informative than the usual spot hybridization technique. Five of the 31 sera were positive for HBV DNA. Levels ranged between 1.36 pq and 143.18 pq per ml of serum, and the levels of HBsAg, anti-HBs, anti-HBc, HBeAg and anti-HBe in the serum were serologically determined. All five sera positive for HBV DNA were also positive for HBsAg. Three of the five positive for HBV DNA were positive for HBeAg and negative for anti-HBe. Two of the sera positive for HBV DNA were negative for HBeAg but positive for anti-HBe. All sera negative for HBV DNA were also negative for HBeAg. Many sera which were negative for HBV DNA and HBeAg were positive for HBsAg. Of the 31 sera from PHC patients, 23 had at least one HBV marker positive (74.2%).     

Prognosis for the patients with chronic hepatitis B

The purpose of the research was to determine the influence of the hepatitis B virus on the progression of the chronic liver disease. In the present paper, 127 patients who were followed up for five years and who had histologically verified chronic liver disease, are described. Fifty two of them were carriers of HBsAg, 75 patients were HBsAg negative, but had other markers typical for a previous infection of HBV in the sera. All the patients were nonalcoholics and no drug addicts. In the sera of these 127 patients markers of HBV were prospectively followed up: HBsAg, HBeAg, anti-HBs, anti-HBc, anti-HBe, HBVDNA, antiHCV for C virus and anti-D for D virus. It was proved by these investigations that HBV provokes very severe chronic hepatitis: CAH (chronic active hepatitis) and CH (cirrhosis hepatis). It was also proved that HBV replicated in 44.20% patients, namely, HBVDNA was positive in the sera of those patients. In 26.08% of such patients the mutant form of HBV was present. In spite of progressive liver disease and without any antiviral therapy all the patients with chronic HBV cirrhosis hepatis were, after five year-follow-up, in Child-Pugh A grade. It was found that the patients who were HBsAg negative, but had one or more markers of HBV positive in the sera, had also a severe chronic hepatitis. That group of patients remains our object of further research. The five-years follow-up of all these patients demonstrates that it is necessary to find out an efficient medicament against HBV chronic hepatitis. Obligatory vaccination of the risk population against virus B remains the only prevention against this severe disease.     

Hepatitis B Virus Genotype Distribution and Genotype-Specific BCP/preCore Substitutions in Acute and Chronic Infections in Argentina

Aim

In order to assess Hepatitis B Virus genotype (g) and subgenotype (sg) implications in the course of infection, 234 HBsAg positive patients in different infection stages were characterized (66 acute infections, 63 HBeAg positive chronic infections and 105 anti-HBe positive chronic infections).

Results

Overall, sgA2 (17.9%), gD (20.9%), sgF1b (34.2%) and sgF4 (19.7%) were the most prevalent. Subgenotype F1b was overrepresented in acute and chronic HBeAg infections (56.1%), whereas gD was the most frequent (40.0%) in anti-HBe positive chronic infections. Among chronic infections, HBeAg positivity rates were 50.0, 12.5, 62.8 and 35.3% for sgA2, gD, sgF1b and sgF4, respectively (p <0.05). A bias toward BCP/preCore mutations was observed among genotypes. In anti-HBe positive chronic infections, sgF1b was more prone to have A1762T/G1764A mutation than sgA2, sgF4 and gD (75.0, 40.0, 33.3 and 31.8%, p<0.005), whereas in the pC region, gD and sgF4 were more likely to have G1896A than sgA2 and sgF1b (81.0, 72.7, 0.0 and 31.3%, p <0.001). The unexpected low frequency of the G1896A mutation in the sgF1b (despite carrying 1858T) prompted us to perform a further analysis in order to identify genotype-specific features that could justify the pattern mutations observed. A region encompassing nucleotides 1720 to 1920 showed the higher dissimilarity between sgF1b and sgF4. Genotypes and subgenotypes carrying the 1727G, 1740C and 1773T polymorphisms were prevented to mutate position 1896.

Discussion

HBeAg seroconversion is a critical event in the natural history of HBV infection. Differences in the HBeAg positivity rate might be relevant since different studies have observed that delayed HBeAg seroconversion is associated with a more severe clinical course of infection, highlighting the critical role that genotypes/subgenotypes might play in the progression of HBV infection. Polymorphisms in the regions 1720 to 1920 could be involved in the molecular mechanisms underlying seroconversion of each genotype/subgenotype.     

Age-specific prevalence of hepatitis B e antigen (HBeAg) and antibody to HBeAg (anti-HBe) among asymptomatic hepatitis B surface antigen (HBsAg) carriers in Okinawa and Kyushu, Japan

A total of 1,741 asymptomatic hepatitis B surface antigen (HBsAg) carriers in two areas (Okinawa and Kyushu) in Japan were surveyed for the presence of hepatitis B e antigen (HBeAg) and the corresponding antibody (anti-HBe) to determine the age-specific prevalence of these markers and the mean age of carriers with HBeAg. Prevalence of HBeAg was significantly higher in Kyushu (36.4% of 755 carriers) than in Okinawa (20.0% of 986 carriers) (P less than 0.001). The mean age of carriers with HBeAg was 25.5 years in Kyushu and 16.1 years in Okinawa, suggesting that HBeAg converted to anti-HBe earlier in Okinawa than in Kyushu. In contrast, the prevalence of anti-HBe was significantly higher in Okinawa (74.6% of 986) than in Kyushu (56.3% of 755) (P less than 0.001). The prevalence of HBeAg decreased with age up to 40-49 years of age and then increased in both areas. Prevalence of anti-HBe was inversely related to the prevalence of HBeAg in both areas. These data suggest that HBeAg and anti-HBe are chronological markers of chronic hepatitis B virus infection and that the duration of HBeAg persistence can be different in different area, even in the same country.     

Low frequency of precore mutants in anti-hepatitis B e antigen positive subjects with chronic hepatitis B virus infection in Chennai, Southern India

The natural course of chronic hepatitis B (CH-B) virus infection is reportedly variable, and the long-term outcomes in hepatitis B e antigen (HBeAg)-negative chronic hepatitis B infection are distinct from HBeAg-positive chronic hepatitis. However, the molecular virological factors that contribute to the progression of liver disease in the south Indian setting remain largely unclear. We prospectively studied 679 consecutive patients for HBsAg, HBeAg, anti-HBeAg, and HBV DNA by qualitative PCR. Randomly selected samples were subjected to bidirectional sequencing to reveal core/precore variants. Of the total 679 chronic HBV cases investigated, 23% (154/679) were replicative HBV carriers. Furthermore, amongst the 560 HBV DNA samples analyzed, 26% (146/560) were viremic. Among the 154 HBeAg positive cases, HBV DNA was positive in 118 cases (77%), significantly (p<0.001) higher than the anti-HBe positive (7%) (28/406) cases. Significant increase in liver disease (p<0.01) with ALT enzyme elevation (p<0.001) was observed in both HBe and anti-HBe viremic cases. Interestingly, low frequencies of mutations were seen in the precore region of the HBV strains studied. HBV precore and core promoter variants were less often detected in subjects with "e" negative chronic HBV infection and, therefore, may not have a prognostic role in determining liver disease sequelae in this part of tropical India.     

Sex- and age-specific prevalences of HBeAg and anti-HBe among HBsAg carriers with or without liver function abnormalities in Okinawa, Japan

We investigated sex- and age-specific prevalences of hepatitis B e antigen (HBeAg) and antibody to HBeAg (anti-HBe) in 1,163 carriers of hepatitis B surface antigen (HBsAg) in Okinawa, Japan, and followed them up for longer than one year in correlation with liver function abnormalities. Elevated serum transaminase levels were found in 160 (13.8%) of them. The prevalence of liver function abnormalities was significantly higher in male carriers (127/690, 18.4%) than in female carriers (33/473, 7.0%) at a P value of 0.001. In asymptomatic carriers, the prevalence of HBeAg was 13.2% and that of anti-HBe 80.0%, significantly different from 41.9% and 54.4%, respectively, in carriers with elevated transaminase levels (P less than 0.001). In asymptomatic carriers, the mean age of HBeAg-positive carriers was 16.7 years which was much lower than the 22.8 years in carriers with liver function abnormalities. The prevalence of elevated transaminase levels was significantly higher in carriers with HBeAg than in those with anti-HBe (33.8% vs. 9.7%, P less than 0.001). Based on these results, the prolonged positivity for serum HBeAg would qualify as a predictor for deteriorating liver function among HBsAg carriers.