The change of the quantitative HBsAg level during the natural course of chronic hepatitis B

Background: There is insufficient information about HBsAg levels and their correlation with serum hepatitis B virus (HBV) DNA in chronic hepatitis B (CHB). Aims: We aimed to describe HBsAg levels during various phases of CHB and to investigate the correlation with serum HBV DNA levels. Methods: A total of 645 treatment‐naïve Korean CHB patients were included in this retrospective cross‐sectional study. They were categorized into immune tolerance (IT, n=56), HBeAg‐positive hepatitis (EPH, n=150), inactive carrier (IC, n=274) and HBeAg‐negative hepatitis (ENH, n=165). The baseline HBsAg and HBV DNA levels were measured. Results: The mean HBsAg titres (log IU/ml) differed (P<0.001): IT 4.29, EPH 3.64, IC 2.05 and ENH 3.23. In 645 patients, HBsAg and HBV DNA showed a significant correlation (r=0.693, P<0.001), and this was also observed in the IT, EPH and IC groups (r=0.664, r=0.541, r=0.505, respectively, all P<0.001), but not in the ENH group (r=0.093, P=0.321). Age had a negative correlation with HBsAg (r=?0.451, P<0.001). The cirrhotic patients had a significantly lower HBsAg level than the non‐cirrhotic patients (2.41 ± 1.36 vs. 3.02 ± 1.21 log IU/ml, P<0.001). Conclusions: The HBsAg level varied significantly in different phases of CHB and was correlated with HBV DNA during the IT, EPH and IC phases. These findings can provide additional information to understand the natural course and pathogenesis of CHB.     

不同临床时期乙型肝炎病毒感染者血清病毒核糖核酸水平的比较

目的观察和分析慢性HBV感染者不同临床时期的血清乙型肝炎病毒核糖核酸(HBV RNA)、HBV DNA以及HBsAg水平。方法入组86例慢性HBV感染者,抽提血清HBV RNA并以Oligo(dT)为引物反转录mRNA为cDNA,Taqmen荧光定量PCR法检测cDNA。将慢性HBV感染者不同临床时期的测定结果进行统计分析。结果血清HBV RNA定量在免疫耐受期、免疫清除期、再活动期、非活动或低(非)复制期患者中依次降低,不同临床时期血清HBV RNA水平两两比较均差异有统计学意义(P0.05)。在慢性HBV感染者中,血清HBV RNA与HBV DNA呈正相关(HBeAg阳性,r=0.523,P=0.004;HBeAg阴性,r=0.340,P=0.034)。血清HBV RNA水平与HBsAg水平均呈正相关(HBeAg阳性,r=0.548,P0.01;HBeAg阴性,r=0.317,P=0.049)。结论随着慢性HBV感染病程的进展,血清HBV RNA水平逐渐下降。且血清HBV RNA水平与HBV DNA、HBsAg水平呈正相关。     

Occult hepatitis B virus infection in Greek patients with chronic hepatitis C and in patients with diverse nonviral hepatic diseases

Occult hepatitis B virus (HBV) infection has been reported in patients with chronic hepatitis C who are negative for HBV surface antigen (HBsAg). However, the significance of 'silent' HBV in hepatitis C virus (HCV) infection is unknown. We investigated 540 subjects for the presence of occult HBV in Greek HCV patients, patients with nonviral liver diseases and healthy donors in an attempt to determine the frequency and importance of this phenomenon. One hundred and eighty-seven anti-HCV(+)/HBsAg(-) patients' sera were investigated for the presence of HBV-DNA by polymerase chain reaction. Two hundred and eighty-two selected blood donors (positive for antibodies to HBV core antigen) and 71 patients with various nonviral hepatic diseases consisted the control groups [both controls were anti-HCV(-)/HBsAg(-)]. HBV-DNA was detected in 26.2% of HCV-infected patients vs 8.5% of patients with nonviral diseases (P = 0.003) and 0/282 of donors (P = 0.0000). HBV-DNA was neither associated with HBV markers, nor with the clinical status of HCV and nonHCV patients. Neither epidemiological, histologic and virologic data nor the response to therapy were associated with the HBV-DNA detection. Hence one quarter of HCV-infected patients had occult HBV infection. Similar findings were not found in both control groups. Occult HBV infection in Greek patients with chronic hepatitis C does not seem to modify the progression of chronic liver disease. Further studies of longer duration are needed in order to clarify the role of 'silent' HBV infection in HCV-infected patients.     

Occult hepatitis B virus infection and its clinical implications

Occult hepatitis B virus (HBV) infection is characterized by presence of HBV infection with undetectable hepatitis B surface antigen (HBsAg). Serum HBV level is usually less than 10⁴ copies/mL in these patients. Diagnosis of occult HBV infection requires sensitive HBV-DNA PCR assay. Several possibilities have been hypothesized as the mechanisms of occult HBV infection. These include: (i) mutations of HBV-DNA sequence; (ii) integration of HBV-DNA into host's chromosomes; (iii) infection of peripheral blood mononuclear cells by HBV; (iv) formation of HBV-containing immune complex; (v) altered host immune response; and (vi) interference of HBV by other viruses. The precise prevalence of occult HBV infection remains to be defined. The clinical implications of occult HBV infection involve different clinical aspects. First of all, occult HBV infection harbours potential risk of HBV transmission through blood transfusion, haemodialysis, and organ transplantation. Second, it may serve as the cause of cryptogenic liver disease, contribute to acute exacerbation of chronic hepatitis B, or even fulminant hepatitis. Third, it is associated with development of hepatocellular carcinoma. Fourth, it may affect disease progression and treatment response of chronic hepatitis C. Most of the previous studies utilized retrospective observation without control groups, and lacked direct association of occult HBV infection with specific pathological changes and disease progression. Highly sensitive, quantitative, and functional molecular analyses of HBV, combined with a well-designed prospective clinical assessment will provide the best approach for the future study of occult HBV infection.     

HBsAg阴性的慢性乙型和丙型重叠感染肝炎患者临床研究

为探讨HBsAg阴性的慢性乙型重叠丙型肝炎患者HBV -DNA水平及临床特征 ,采用PCR -微板核酸杂交 -ELISA法对 2 0 3例HBVM一项 /一项以上阳性的慢性丙型肝炎患者进行HBV -DNA定量及HCV -RNA基因分型检测 ,并分析患者性别、年龄以及肝功能等临床特征。HBsAg阴性及HBsAg阳性慢性丙型肝炎患者在性别、年龄分布上无差异 ;但血清ALT及总胆红素水平在HBsAg阳性者较高。Ⅱ / 1b型HCV感染在HBsAg阴性者中的比例高于HBsAg阳性者 (P <0 0 5 )。HBsAg阴性慢性丙肝患者HBV -DNA阳性率及HBV水平均低于HBsAg阳性患者。HBsAg阴性的慢性丙肝患者中 ,抗HBc单独阳性者HBV -DNA阳性率最高 ,但HBV -DNA水平无显著性差异。Ⅱ /1b型HCV感染在HBV -DNA阴性患者中的出现频率较高 ,与HBV -DNA阳性患者相比差异有显著性。因此 ,临床上对于那些肝功能损害严重 ,同时有抗HBc阳性或Ⅱ / 1b型HCV感染 ,尤其是HCV -RNA阴性而肝功能仍有持续损害的患者要及时行HBV -DNA检测 ,以除外HBV活动的可能     

Correlation between HBsAg quantitative assay results and HBV DNA levels in chronic HBV

Background

Viral load has been used to diagnose and monitor patients who are being treated for chronic hepatitis B (CHB). The Diagnosis methods are molecular-based and expensive. Quantitation of hepatitis B surface antigen (HBsAg) by automated chemiluminescent micro-particle immunoassay has been proposed to be a surrogate marker. Quantitating HBV DNA levels molecularly is expensive; thus, a cheaper laboratory test as a surrogate diagnostic marker might simplify our management.

Objectives

We determined whether quantitative HBsAg levels correlate with HBV DNA levels in CHB.

Patients and Methods

In this cross-sectional study, all CHB patients who were referred by a gastroenterologist to undergo quantitative HBV DNA assay in a qualified laboratory in Mashhad, Iran in 2009 were enrolled, and blood samples was obtained. Patients who were positive for antibodies to HCV and HDV were excluded. HBV DNA was measured by real-time polymerase chain reaction, and serum HBsAg was quantified byelectrochemiluminescence assay (Roche Diagnostic).

Results

Of 97 patients, 70 were male (72%) and 27 were female (28%); the mean age was 39 ± 11 years. Eighty-seven percent wasHBeAg-negative. By Mann-Whitney test,HBSAg titer differed significantly between HBeAg-positive and -negative patients (P = 0.001), as did HBV DNA levels (P = 0.009). By Spearman test, there was no significant correlation between HBsAg and HBV DNA levels (P= 0.606 and r = 0.53).

Conclusions

HBeAg-negative patients have higher levels of HBsAg and lower levels of HBV DNA. By electrochemiluminescence assay,HBsAg has no significant correlation with HBV DNA levels in CHB with predominant genotype D and HBeAg negativity in Iran.     

慢性乙型肝炎患者肝组织中HBV cccDNA定量与病情的相关性分析

目的探讨肝组织HBV cccDNA定量对慢性乙型肝炎（乙肝）患者病情的影响。方法应用FQ—PRC法检测42例乙肝患者肝组织HBVcecDNA、肝组织和血清HBV—DNA水平,同时对肝组织行常规病理染色判断肝组织炎症及纤维化程度;SP法检测肝组织中HBsAg、HBcAg表达,化学发光法检测HBV标志物。分析肝组织HBVceeDNA与组织和血清HBV-DNA、肝细胞内HBsAg、HBcAg水平及肝脏炎症、纤维化程度的关系。结果肝组织HBVeecDNA定量与组织及血清HBV—DNA定量呈正相关（r=0．807,P〈0．001;r=0．627,P〈0．001）;与肝组织炎症活动度及纤维化程度无明显相关性：肝组织HBVcccDNA定量与肝细胞内HBsAg表达无明显相关性,而与HB—cAg表达呈正相关（r=0．486,P〈0．05）。结论检测肝组织内HBVceeDNA可更精确反映HBV复制程度;对乙肝诊断、抗病毒治疗及选择停药时机具有重要价值。肝组织HBV cccDNA与血清HBV-DNA定量及HBcAg联合检测可指导抗病毒治疗。     

Correlation of hepatitis B virus (HBV) genotypes and mutations in basal core promoter/precore with clinical features of chronic HBV infection.

BACKGROUND/AIMS: To investigate the correlation of hepatitis B virus (HBV) genotypes and basal core promoter (BCP) and precore (PC) mutations in patients with chronic hepatitis B. METHODS: HBV genotyping, nucleotide mutation, serum HBV DNA level and serological markers were analyzed in 121 patients with chronic HBV infection using INNO-LiPA HBV genotyping, polymerase chain reaction (PCR) product-based sequencing, fluorescence quantitative PCR and enzyme-linked immunosorbent assays respectively. RESULTS: Forty (33.0%), 77 (63.6%), two (1.7%) and two (1.7%) patients had genotypes B, C, B/C and D infections respectively. Significant differences were found in serum HBV DNA levels (log10 copies/ml: 6.18 vs. 5.61, P=0.042) and mutations at nucleotide (nt) 1762/1764 (71.4% vs. 42.5%, P=0.002) between genotypes C- and B-infected patients. There were significant differences in the mean age, serum biochemical parameter levels and mutation rates in BCP/PC among hepatitis e antigen (HBeAg)-positive and -negative chronic hepatitis B (CHB) and liver cirrhosis (LC) groups. CONCLUSION: Genotypes C and B are predominant in China, and the frequent nt 1762/1764 mutation, which occurs commonly in HBeAg-negative CHB, especially in genotype C patients, may be associated with the progress of chronic HBV infection.     

Effects of interferon-α therapy on serum and liver HBV DNA in patients with chronic hepatitis B

The aim of this study was to evaluate the effect of interferon- therapy on serum and liver HBV DNA in 20 patients with chronic hepatitis B and to correlate the presence or absence of HBV DNA with the clinical response. There were 11 responders and all lost HBV DNA from the serum. Ten of the 11 were followed for 36 months following IFN treatment and remained well with absence of HB_eAg and HBV DNA from the serum and with normal ALT. Five also lost HB_sAg. HBV DNA became undetectable in the liver of nine of 10 of these patients in whom liver tissue was available for study. HBV DNA persisted in the liver of seven of nine nonresponders and was not detected in two in spite of the presence of HBV DNA and HB_eAg in the serum of these two patients. We conclude that IFN may induce long remissions in patients with chronic hepatitis B with loss of HBV DNA from the serum and that occasionally HBV DNA may persist in the liver of such patients.This work was supported in part by Health and Welfare Canada Grant 6606-2435-52.     

Hepatitis B Virus Genomes of Chronic Hepatitis Patients Do Not Contain Specific Mutations Related to Acute Exacerbation

To determine the specific viral variants associated with acute exacerbation of chronic hepatitis from hepatitis B virus (HBV) infection, we analyzed the complete nucleotide sequences of the HBV genome in serial serum samples from two chronic active hepatitis patients who seroconverted from HBeAg to anti-HBe. HBV DNA was amplified by polymerase chain reaction (PCR) and sequenced. A 1896 precore stop codon mutant (G to A at nt 1896) coexisting with the wild sequence was found in both patients prior to seroconversion from HBeAg to anti-HBe. Core promoter mutations at nucleotide positions 1762 (A to T) and 1764 (G to A) were found in both patients throughout the observation period. Mutations were observed in the HBV genome of the two patients at different time points, and there was no correlation between the mutations and liver disease or DNA polymerase levels. The nucleotide divergence rate and the composition of quasispecies in the HBV sequence at the time of acute exacerbation were almost the same as were found at other time points. These results suggest that acute exacerbation does not appear to be caused by a characteristic HBV species. The multiple factors that cause generalized HBV replication activation may contribute to acute exacerbation.     

Hepatitis B virus (HBV) DNA levels and the management of HBV-infected health care workers

Different guidelines exist for the management of hepatitis B virus (HBV)-infected health care workers (HCWs). Various HBV DNA levels are used as a cutoff level to determine whether an HBV-infected HCW is allowed to perform exposure-prone procedures (EPPs) or not. In this paper we discuss the factors that determine HBV DNA levels and the implications of different HBV DNA cutoff levels for EPP performing HCWs. If the level of HBV DNA in the serum of HCWs is used to determine acceptability for the conduct of EPPs, it is necessary to take into account the variability in time of HBV DNA levels in HBV carriers and the reliability and reproducibility of the molecular diagnostic test involved. The issue of standardization has to be addressed, before a universal, maximum level of viraemia for EPP performing HCWs can be introduced.     

Hepatitis B virus genotypes and G1896A precore mutation in 486 Spanish patients with acute and chronic HBV infection

This study aims to determine the prevalence of hepatitis B virus (HBV) genotypes (A-F) and their association with the G1896A precore mutation in 486 patients positive for HBV surface antigen. Genotypes were determined by RFLP and precore mutation by real-time PCR. Genotypes D (48.1%) and A (39.5%) were the most common, followed by F (4.1%) and B, C and E (<1%). The A to D ratio (A:D) was 1.4 in HBeAg+ chronic hepatitis B (CHB), 0.6 in HBeAg- CHB and 1.4 in HBeAg- inactive carriers. Distribution of these genotypes was different between HBeAg+ CHB and HBeAg- CHB (P = 0.02), and between HBeAg- CHB and HBeAg- inactive carriers (P = 0.009). Genotype A was the most prevalent in HBeAg+ CHB with elevated alanine aminotransferase (ALT) (68.6%) and genotype D in HBeAg+ CHB with fluctuating ALT (60.7%). There was a difference in genotype prevalence between chronic and acute infection (P = 0.03). The precore mutant correlated with high levels of HBV-DNA in genotype d HBeAg- CHB. Genotype D is not as highly prevalent in Spanish patients as would be expected in a Mediterranean area. The unequal prevalence of genotypes between acute and chronic infection suggests that genotype A is associated with a higher tendency to cause chronic infection.     

HBV感染血清标志物、HBV DNA水平与性别的关系

探讨HBV感染者血清标志物、HBVDNA水平和性别之间的关系.选择495例HBsAg阳性的血清标本和502例HBV DNA高于1×108copy/ml以上的血清标本,分别通过ELISA法以及实时荧光定量法进行HBV感染血清标志物HBV DNA的检测.三种组合[组合1为HBsAg( )HBeAg( )抗-HBc( ),组合2为HBsAg( )抗-HBe( )抗-HBc( ),组合3为HBsAg( )抗-HBc( )]的HBV DNA阳性率(分别为95%、18%和53%)存在极显著差异(P＜0.01);女性在HBAg阴性模式中的比例与在HBeAg阳性模式中的比例差异显著(P＜0.05).HBV感染血清标志物、HBV DNA水平和性别之间存在着重要的关系;对于组合3的患者要给予足够重视;女性在高HBV DNA水平而且HBeAg阴性组合中的比例非常小,这也许与女性在肝癌患者中所占比例较低存在着一定的关系.     

乙型肝炎患者检测肝组织HBV-DNA的意义

为了探讨提高乙型肝炎病毒微量检出率的有效途径。采用PCR分别检测血清及肝穿刺组织HBVD-NA，血清检出总阳性率为41.7%，肝组织检出总阳性率为72.9%。在血清HBsAg阴性的病例中PCR血清检出阳性率为26.7%。肝组织检出阳性率为66.7%。早此可见PCR检测灵敏。准确、特别对血清HBVM难以检测的微量HBVDNA更有价值。两种标本来源相比肝组织较血清为优。