HBV感染者基因型、HBVDNA、HBV cccDNA、ALT血清水平变化与抗病毒治疗相关性分析

为检测HBV感染者基因型及血清HBV DNA、HBV cccDNA、ALT、HBeAg水平,探讨它们之间相互关系及治疗后与这些指标的相关性,采用PCR荧光定量、基因测序、化学发光、生化分析等方法,分别对收集的202例HBV感染的患者Lamivudine抗病毒治疗前后的血清HBV DNA、HBV cccDNA、 ALT、 HBeAg定量检测及HBV DNA基因分型.结果显示,HBV B型和C型患者的血清 HBV DNA和cccDNA的水平没有显著性差异,HBV cccDNA与HBV DNA的比值也没有显著性差异;HBV B型和C型患者的血清 HBV DNA和cccDNA的水平与ALT有相关性;不管是HBV B型还是C型的患者,HBeAg阳性的患者血清中HBV DNA和cccDNA显著高于HBeAg阴性的患者;但是,C型的患者,HBeAg阳性的患者cccDNA与HBV DNA的比值显著低于HBeAg阴性的患者;治疗后比治疗前除 HBeAg没有显著性差异(P>0.05),HBV DNA、cccDNA、ALT均有显著性差异(P<0.01).感染HBV B型和C型的患者血清 HBV DNA和cccDNA的水平无显著性差异;而C型的患者,HBeAg阳性的患者cccDNA与HBV DNA的比值显著低于HBeAg阴性的患者;HBV DNA基因型、HBV cccDNA、ALT与抗病毒治疗密切相关.     

乙肝患者病毒核酸载量与乙肝病毒外膜大蛋白含量相关性的研究

目的：探讨乙型肝炎患者病毒核酸载量与血清乙型肝炎病毒外膜大蛋白（HBV-LP）含量的关系。方法：采用酶联免疫吸附试验（ELISA）和实时荧光定量PCR法分别检测307例乙型肝炎患者血清的HBV-LP、HBV-M和HBV DNA,并进行相关性分析。结果：307例乙型肝炎患者血清中HBV-LP含量与HBV DNA拷贝数变化相一致,两者呈正相关（r=0.943）;在131份HBeAg阳性血清中,HBV DNA与HBV-LP的阳性率分别为95.42%和93.89%,两者差异无显著性（P〉0.05）;在176份HBeAg阴性血清中,HBV DNA与HBV-LP的阳性率分别为46.02%和56.25%,两者差异无显著性（P〉0.05）。结论：血清HBV-LP水平能反映HBV感染者体内HBV复制程度,其灵敏度高于HBeAg,可作为判断HBV复制新的血清学指标。     

慢性乙肝患者血清趋化因子RANTES水平与肝功能生化指标等的相关性研究

目的 了解慢性乙型肝炎(慢性乙肝)患者血清趋化因子RANTES水平,探讨血清RANTES水平与丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、总胆红素(TBil)、凝血酶原活动度(PTA)、乙型肝炎e抗原(HBeAg)及乙型肝炎病毒(HBV DNA)载量的相关性.方法 选择144例慢性乙肝患者(观察组)和18名健康人(对照组),采取静脉血并应用ABC-ELISA方法 检测其血清中趋化因子RANTES浓度,并与两组的肝功能检测生化指标、HBeAg和HBV DNA载量进行相关性分析,利用SPSS13.0软件进行统计分析.结果 慢性乙肝患者血清RANTES的浓度比正常对照组升高,血清RANTES浓度分别为(3930.12±2856.96)ng/ml和(329.46±152.23)ng/ml,两组之间差异比较均有统计学意义(P<0.05);RANTES水平与ALT(r=0.197,P:0.018)、AST(r=0.239,P=0.004)和Tnil(r=0.316,P=0.001)呈显著正相关;RANTES水平与PTA(r=-0.078,P=0.357)无显著相关;HBeAg阴性组与HBeAg阳性RANTES水平比较无统计学意义(P=0.407);HBV DNA低载量组(<105拷贝/ml)和HBVDNA高载量组(≥105拷贝/ml)RANTES水平比较无统计学意义(P=0.185).结论 慢性乙肝患者血清中RANTES表达水平增高,血清RANTES水平与ALT、AST和TBIL呈正相关,与PTA无相关性.RANTES水平可反映肝脏炎症活动及损害情况,不受HBeAg、HBV DNA载量影响,可能参与慢性乙肝发病.     

HBeAg阴性慢性乙型肝炎患者血清乙型肝炎病毒外膜大蛋白检测分析

目的 探讨血清乙型肝炎病毒外膜大蛋白(HBV-LP)检测在HBeAg阴性慢性乙型肝炎中的诊断价值和临床意义.方法 分别采用酶联免疫吸附试验(ELISA)法和实时荧光定量PCR法检测306份慢性乙型肝炎患者血清中HBV.M、HBV-LP和HBV.DNA.结果 (1)在未经过抗病毒治疗的患者中,HBV-LP阳性率和HBV DNA无明显差异,而且HBV-LP的吸光度(A值)和HBV DNA拷贝数呈正相关性;(2)在经过抗病毒治疗的患者中HBV-LP的阳性率明显高于HBV DNA.结论 血清中乙型肝炎病毒大蛋白水平能反映HBV感染者体内HBV复制程度,对于判断HBeAg阴性患者体内复制具有重要的临床意义.     

高灵敏度乙肝核酸定量与乙肝血清标志物的相关性分析

目的 探讨高灵敏度的荧光定量PCR检测的乙型肝炎病毒核酸(HBV DNA)与乙肝血清免疫标志物(HBV M)之间的相关性及临床意义.方法 选取234例患者标本,同时检测HBV DNA载量和HBV M.根据HBV M模式将结果分为五组,并对HBV DNA载量和HBV M之间的关系进行分析.选取177例乙肝患者,分析HBV DNA载量水平与HBeAg之间的关系.结果 Ⅰ组HBV DNA阳性率为77.4％;Ⅱ组HBV DNA阳性率为70.4％,两者之间差异无统计学意义(χ2=0.617,P ＞0.05).HBeAg表达水平与HBV DNA载量之间存在正相关关系(γ=0.812,P＜0.01).结论 高灵敏度HBVDNA与经典化学发光法的检测结果之间有很好的相关性.提高HBV DNA的检测灵敏度对于乙型肝炎患者,尤其对小三阳患者的病情和疗效评估具有重要的临床意义.     

慢性乙型肝炎患者肝脏病理特点与血清HBeAg和HBV DNA的关系

目的 了解慢性乙型肝炎患者病理特点与血清HBeAg和HBVDNA的关系.方法对1057例慢性乙型肝炎患者进行肝脏病理检查,采用荧光定量PCR法检测血清HBV DNA,用化学发光法检测血清HBeAg.结果 HBeAg阴性的慢性乙型肝炎患者的炎症及纤维化程度(G4和S4分别为7.83%和12.17%)较HBeAg阳性的慢性乙型肝炎患者高(G4和S4分别为3.39%和5.44%);HBeAg阳性的患者中HBV DNA滴度低的患者炎症及纤维化程度较高(HBV DNA 104～105 G3G4和S3S4分别为45.64%和30.20%),而HBeAg阴性的患者则是HBV DNA滴度高的炎症及纤维化程度较高(HBV DNA106～107 G3G4为54.55%和HBV DNA 108～109S3S4为42.85%).结论慢性乙型肝炎患者的肝脏病理与血清HBeAg及HBV DNA水平有不同相关性,对HBeAg阴性的慢性乙型肝炎患者要及早进行肝脏病理检查和抗病毒治疗.     

HBV感染者基因、HBV DNA、HBV cccDNA、ALT血清水平变化与抗病毒治疗相关性分析

目的：为检测HBV感染者基因型及血清 HBV cccDNA、HBV DNA、ALT、HBeAg水平,探讨它们之间相互关系及治疗后与这些指标的相关性,采用PCR荧光定量、基因测序、化学发光、生化分析等方法,分别对收集的202名HBV感染的患者治疗前后（经过使用抗病毒药物---拉米呋定,一个疗程）的血清HBV DNA、 cccDNA、 ALT、 HBeAg定量检测及HBV DNA基因分型。结果显示,HBV B型和C型患者的血清 HBV DNA和cccDNA的水平没有显著性差异,cccDNA与HBV DNA的比值也没有显著性差异;HBV B型和C型患者的血清 HBV DNA和cccDNA的水平与ALT有相关性;不管是HBV B型还是C型的患者,HBeAg阳性的病人血清中HBV DNA和cccDNA显著高于HBeAg阴性的病人;但是,C型的患者,HBeAg阳性的病人cccDNA与HBV DNA的比值显著低于HBeAg阴性的病人;治疗后比治疗前除 HBeAg没有显著性差异（P ＞0.05）,HBV DNA、 cccDNA、 ALT均有显著性差异（P <0.01）。结论：感染HBV B型和C型的患者血清 HBV DNA和cccDNA的水平无显著性差异;而C型的患者,HBeAg阳性的病人cccDNA与HBV DNA的比值显著低于HBeAg阴性的病人;HBV基因型、HBV cccDNA、ALT与抗病毒治疗密切相关。     

孕妇乙型肝炎HBV-M与preS1-Ag、HBV DNA的相关性及联合检测的临床价值

目的分析乙型肝炎孕妇HBV血清标志物（HBV-M）、HBV前S1抗原（preS1-Ag）及HBV DNA的相关性,探讨联合检测的临床价值。方法采用化学发光免疫分析法对375例乙型肝炎孕妇血清进行HBV-M的检测,用ELISA法检测preS1-Ag,同时采用荧光定量PCR法检测HBV DNA。结果在不同模式乙型肝炎孕妇血清中,preS1-Ag、HBeAg的检出率分别是61.87%、34.67%;在HBeAg阳性血清中,preS1-Ag检出率为90%,HBeAg阴性血清的检出率为46.94%,两者之间有统计学意义;HBV DNA与HBeAg一致性比较较差,而与preS1-Ag一致性比较较好。结论 HBVpreS1-Ag较HBeAg更敏感地反映了HBV的复制情况,HBV preS1-Ag和HBV DNA有较高的符合率,可作为HBV复制的指标,三者联合检测,可及时阻断乙肝母婴传播,降低婴儿乙肝感染率。     

e抗原阳性慢性乙型肝炎患者血清HBV-DNA与HBsAg、HBeAg的相关性分析

目的 研究e抗原阳性慢性乙型肝炎患者外周血中HBV-DNA载量与乙型肝炎病毒表面抗原（HBsAg）、乙型肝炎病毒e抗原（HBeAg）的相关性,及其在不同性别、年龄群体中的差异.方法 收集319例e抗原阳性慢性乙肝患者血清,采用实时荧光定量PCR法检测HBV-DNA载量,用时间分辨免疫荧光法检测HBsAg和HBeAg的浓度,利用SPSS软件做统计分析.结果 HBV-DNA载量与HBsAg含量有良好的相关性（r=0.514,P〈0.001）;与HBeAg含量有相关（r=0.337,P〈0.001）;女性的HBeAg水平要高于男性患者（P〈0.05）;年龄（31～50）岁组、〉50岁组的HBV-DNA、HBsAg 及HBeAg值皆高于年龄 〈30岁组 （P〈0.001）.结论 e抗原阳性慢性乙型肝炎患者血清中HBV-DNA载量与HBsAg、HBeAg定量水平皆有相关性,其中与HBsAg相关性更佳.     

慢性乙型肝炎外周血单个核细胞内HBV-DNA与血清HBV-DNA及HBeAg表达关系的研究

目的研究慢性乙型肝炎患者外周血单个核细胞(PBMC)内HBV-DNA和血清中HBV-DNA表达量、e抗原表达的关系。方法采用聚合酶链反应(PCR)检测208例慢性乙型肝炎患者PBMC内HBV-DNA,应用荧光定量聚合酶链反应(FQ-PCR)检测血清中HBV DNA含量,应用酶联免疫吸附(ELISA)法检测乙肝血清标志物。结果208例慢性乙型肝炎患者PBMC内HBV-DNA阳性106例、阴性102例。HBV-DNA(PBMC)阳性组、阴性组血清HBV-DNA定量≥1.0E5患者比例分别为91.5%(97例)、45.1%(46例)(χ2=52.12,P<0.01);HBeAg阳性率分别为76.4%(81例)、50.9%(52例)(χ2=21.55,P<0.01)。结论PBMC内HBV-DNA的检测与血清中HBV-DNA定量检测及HBeAg阳性率存在明显的正相关。提示血清HBV-DNA高载量的HBeAg阳性患者外周血单个核细胞感染HBV-DNA明显增加。     

Correlation between serum hepatitis B virus core‐related antigen and intrahepatic covalently closed circular DNA in chronic hepatitis B patients

Nucleos(t)ide analogues are utilized for the treatment of chronic HBV infection, and HBe seroconversion and HBV DNA levels are commonly used as markers of viral status and as primary treatment endpoints. Recently, a new assay was prepared for the detection of serum HBV core‐related antigen (HBcrAg), consisting of HBcAg, HBeAg, and p22cr, which is a precore protein from amino acid ?28 to at least amino acid 150, by coding the precore/core region. In this study, we examined the correlation between serum HBcrAg concentration and viral status by the analysis of serum HBeAg, HBsAg, peripheral HBV DNA, and intrahepatic covalently closed circular DNA (cccDNA) in 57 chronic hepatitis B patients. Intrahepatic cccDNA was detected in all 57 patients, 42 patients were HBcrAg‐positive, and serum HBcrAg concentration level was closely correlated with cccDNA. Additionally, positive HBcrAg concentration level results were observed in 6 out of 13 HBsAg seroclearance patients and 20 out of 31 HBV DNA‐negative patients. Moreover, the correlation between HBcrAg and cccDNA in these 31 HBV DNA‐negative patients was statistically significant (r = 0.482, P = 0.006). These data suggest that serum HBcrAg concentration is well correlated with intrahepatic cccDNA level, and that the measurement of serum HBcrAg may be clinically useful for monitoring intrahepatic HBV viral status, especially in patients under treatment with nucleos(t)ide analogues. J. Med. Virol. 81:27–33, 2009. © 2008 Wiley‐Liss, Inc.     

Validation of a sensitive and specific real‐time PCR for detection and quantitation of hepatitis B virus covalently closed circular DNA in plasma of chronic hepatitis B patients

Hepatitis B virus (HBV) covalently closed circular DNA (cccDNA) serves as a template for viral replication and plays a role in persistence of HBV infection. The origin and significance of cccDNA in plasma however, is not well understood. A sensitive, specific, and reproducible real‐time PCR for detection and quantitation of cccDNA in plasma of chronic hepatitis B patients was developed and validated. Four HBV DNA reference panels, and 96 plasma samples of chronic hepatitis B patients were analyzed. Results were compared with total HBV DNA levels, individual ALT levels and the Histology Activity Index (HAI). This cccDNA assay had a lower limit of detection at 15 copies/PCR, a lower limit of quantitation at 91 copies/PCR and a correlation coefficient (R) of 0.98 (P < 0.0001). cccDNA was detected in two of four international panels. Significant correlation was found between cccDNA and total HBV DNA levels in both panels (R = 0.96, and R = 0.43) and in samples of the chronic hepatitis B patients (R = 0.88, P < 0.0001). In 57% of these samples cccDNA was detectable. Mean level of cccDNA was 0.16% of total HBV load. Plasma cccDNA levels were higher in HBeAg positive samples than in HBeAg negative samples (4.91 log copies/ml vs. 3.88 log copies/ml, P < 0.0001). Levels of total HBV DNA and HBV genotype did not influence cccDNA detection. ALT levels and HAI‐score were not correlated with plasma cccDNA levels. These findings suggest that cccDNA levels in plasma are not the result of increased hepatocyte degeneration, but indicate that other mechanisms might be responsible. J. Med. Virol. 81:988–995, 2009. © 2009 Wiley‐Liss, Inc.     

Relationship between serum hepatitis B virus DNA and surface antigen with covalently closed circular DNA in HBeAg‐negative patients

Hepatitis B virus (HBV) covalently closed circular DNA (cccDNA) is responsible for viral persistence. This study aimed to investigate the serum surrogate markers for cccDNA and to evaluate the intrahepatic viral events associated with disease activity in HBeAg‐negative chronic hepatitis B patients. Thirty‐three treatment‐naïve patients with a negative HBeAg who had a liver biopsy were studied. Active disease was defined as a serum alanine aminotransferase >40 IU/L and a serum HBV DNA >10,000 copies/ml. This study showed significant correlation between serum HBV DNA and both log cccDNA (r = 0.41, P = 0.018) and log total intrahepatic HBV DNA (r = 0.71, P < 0.0001). No significant correlation was observed between serum HBsAg and log cccDNA (P = 0.15) or log total intrahepatic HBV DNA (P = 0.97). Fourteen and 19 patients had inactive and active disease, respectively. The median log cccDNA and log total intrahepatic HBV DNA (copies/10⁶ cells) were significantly higher in patients with active disease compared with those with inactive disease (4.11 vs. 3.53, P = 0.03 and 5.46 vs. 4.64, P < 0.001, respectively). The HBV replicative efficiency, defined as the ratio of serum HBV DNA to cccDNA, was approximately 20% higher in patients with active disease. No significant difference was observed in the HBsAg levels and the ratio of serum HBsAg to cccDNA between the two groups. In conclusion, serum HBV DNA, but not HBsAg, reflects the amount of cccDNA and the replication efficiency of HBV in patients with HBeAg‐negative chronic hepatitis B. J. Med. Virol. 82:1494–1500, 2010. © 2010 Wiley‐Liss, Inc.     

Identification of different states of hepatitis B virus infection with a quantitative PCR assay

The level of hepatitis B virus (HBV) DNA in serum reflects the replicative activity of HBV. To compare serum HBV DNA levels in different states of hepatitis B, 47 sera of patients with HBeAg-positive chronic hepatitis B, 4 sera of patients with HBeAg-negative chronic hepatitis B, 40 samples of patients after HBeAg seroconversion during alpha interferon treatment, 57 sera of inactive HBsAg carriers, and 42 sera of patients who had recovered from chronic hepatitis B more than 12 months prior to blood collection were checked for the presence of HBV DNA with the Amplicor HBV Monitor Test. In patients with HBeAg-positive chronic hepatitis B, the median of serum HBV DNA levels (8.3 x 10(8) copies/ml) was significantly higher than that for patients after HBeAg seroconversion (6.2 x 10(3) copies/ml) and than that for inactive HBsAg carriers (5.6 x 10(3) copies/ml). None of the patients who had recovered from hepatitis B had detectable HBV DNA in serum. Quantitative PCR proved to be a valuable tool for identification of different states of HBV infection. This technique was found to be a good method for determination of serum HBV DNA levels both for patients with HBeAg seroconversion and for inactive carriers who showed low viremia not detectable by conventional hybridization assays.     

Hepatitis B virus core-related antigens as markers for monitoring chronic hepatitis B infection

A sensitive chemiluminescence enzyme immunoassay has been developed for hepatitis B virus (HBV) core-related antigen (HBcrAg) detection. We aimed to investigate the usefulness of HBcrAg measurement for monitoring chronic hepatitis B disease. HBcrAg levels were measured by a chemiluminescence enzyme immunoassay in 54 untreated patients and 39 patients treated with either entecavir or lamivudine. The HBcrAg concentration correlated positively with the levels of serum HBV DNA (r = 0.820), intrahepatic total HBV DNA (r = 0.700), and covalently closed circular DNA (cccDNA) (r = 0.664; for all, P values were <0.001). A higher HBcrAg concentration was associated with a greater proportion of hepatitis B core antigen immunostaining. Although the differences were not statistically significant, patients with higher Knodell necroinflammation and fibrosis scores tended to have higher serum HBcrAg concentration levels. In the treated patients, the logarithmic reduction in HBcrAg at week 48 correlated positively with the logarithmic reduction of serum HBV DNA, intrahepatic total HBV DNA, and cccDNA. Of the 31 patients with undetectable serum HBV DNA (<300 copies/ml) at the end of treatment, 20 (65%) still had detectable HBcrAg. A greater reduction in posttreatment HBcrAg concentration was associated with histological improvement and a decrease in hepatitis B core antigen immunostaining. HBcrAg concentrations of <40,000 kU/ml at baseline and <200 kU/ml at week 24 were associated with a higher chance of having undetectable HBV DNA at week 48. In conclusion, serum HBcrAg levels correlated with HBV virological markers and reflected the chronic hepatitis B disease activity in the liver.     

HBVcccDNA在乙型肝炎患者血清、外周单核细胞及肝组织中的分布

目的 观察乙肝病毒共价闭合环状DNA(HBVcccDNA)在乙肝患者血清、外周单核细胞(PBMC)及肝组织中的分布情况.方法 选取血清HBVDNA＞105拷贝/ml的乙肝患者50例,血清HBVDNA＜105拷贝/ml的乙肝患者30例,脂肪肝患者(非乙型肝炎)20例,同时采用实时荧光定量聚合酶链反应检测患者血清、PBMC及肝组织中HBVcccDNA的存在情况.结果 50份血清HBVDNA＞105拷贝/ml的标本中血清HBVcccDNA检出28例,检出率56%,PBMC HBVcccDNA检出29例,检出率58%,肝组织中HBVcccDNA检出44例,检出率88%,血清、PBMC的检出较肝组织检出差异有统计学意义P＜0.005,血清较PBMC检出差异无统计学意义P＞0.005.30份血清HBVDNA＜105拷贝/ml的标本中血清、PBMC HBVcccDNA检出均为2例,检出率6.67%,肝组织中HBVcccDNA检出6例,检出率20%,血清、PBMC、肝组织三者之间检出差异无统计学意义,P＞0.005.20份脂肪肝患者的血清、PBMC及肝组织中均未检出HBVcccDNA.结论 HBVcccDNA主要存在于乙型肝炎患者的肝脏中,乙肝患者血清及PBMC中也有HBVcccDNA的存在但较肝组织中要少的多.     

Serum hepatitis B surface antigen is correlated with intrahepatic total HBV DNA and cccDNA in treatment‐naïve patients with chronic hepatitis B but not in patients with HBV related hepatocellular carcinoma

The aim of the study was to investigate correlations between intrahepatic hepatitis B virus total DNA, covalently closed circular DNA (cccDNA), and serum HBsAg in treatment‐naïve chronic hepatitis B and HBV related hepatocellular carcinoma (HCC). Liver tissues were taken from 42 HBV related HCC and 36 patients with chronic hepatitis B. A fraction of DNA extracted from liver tissue was digested with a plasmid‐safe ATP‐dependent DNase and used for HBV cccDNA detection. The remaining DNA was used for the detection of HBV total DNA and β‐globin, the latter of which is a housekeeping gene and quantified for normalization by real‐time PCR. Quantitation of serum HBsAg was performed by a chemiluminescence assay. Serum HBsAg had positive correlations with serum HBV DNA (r = 0.636, P < 0.001), intrahepatic HBV total DNA (r = 0.519, P = 0.001) and cccDNA (r = 0.733, P < 0.001) in 36 treatment‐naïve chronic hepatitis B, while HBsAg correlated poorly with DNA (r = 0.224, P = 0.210), intrahepatic total DNA and cccDNA in the tumor (r = 0.351, P = 0.031; r = 0.164, P = 0.324, respectively) and non‐tumor (r = 0.237, P = 0.152; r = 0.072, P = 0.667, respectively) liver tissues of 42 HCC. HBV cccDNA and total DNA were significantly higher in liver tissue from chronic hepatitis B than in tumor and non‐tumor of HCC (P < 0.001). Serum HBsAg and HBV DNA were also higher in chronic hepatitis B than in HCC (P < 0.001). It was concluded that levels of serum HBsAg and intrahepatic cccDNA and total DNA were significantly higher in chronic hepatitis B than in HCC, and significant correlations among them were observed in treatment‐naïve chronic hepatitis B but not in HCC. J. Med. Virol. 85:219–227, 2013. © 2012 Wiley Periodicals, Inc.     

Detection and characterization of hepatitis B virus DNA in serum of HBe antigen-negative HBsAg carriers

Sera from 153 Israeli patients in various stages of hepatitis B virus (HBV) infection with undetectable hepatitis Be antigen (HBeAg) were studied for the presence of HBV DNA in the serum by molecular hybridization. HBV DNA was detected in 10 patients: 3 with acute hepatitis, 4 asymptomatic hepatitis B surface antigen (HBsAg) carriers, 1 with chronic active hepatitis, 1 with cirrhosis, and 1 with mixed cryoglobulinemia. HBV DNA was detected in 7 of 10 HBeAg-positive control samples tested. Hybridization analysis was used for quantitative comparison of HBV DNA levels in serum. HBV DNA levels, found in HBeAg-negative patients sometimes exceeded the levels found in HBeAg-positive patients. Restriction enzyme analysis of serum HBV DNA from four HBeAg-negative samples gave undistinguishable digestion patterns as compared to 3 HbeAg-positive samples. However, heterogeneity in HBV DNA restriction fragments was detected among HBV genomes in sera of HBeAg-positive samples. These data demonstrate that HBV DNA may be present in the serum at various stages of HBV infection, regardless of HBeAg detection. Failure to detect HBeAg in these patients does not necessarily reflect low serum levels of viral particles, or the occurrence of HBV genome variants.     

Impact of peginterferon alpha‐2b and entecavir hydrate combination therapy on persistent viral suppression in patients with chronic hepatitis B

The ideal approach to treat chronic hepatitis B remains controversial. This pilot study aimed to evaluate the effectiveness of peginterferon (PEG‐IFN) α‐2b and entecavir hydrate (ETV) as a combination therapy for patients with chronic hepatitis B, particularly in the context of virological response and the reduction of intrahepatic covalently closed circular DNA (cccDNA). A total of 17 patients with hepatitis B virus (HBV) genotype C were enrolled in this study. All subjects were treated with this combination therapy for 48 weeks and observed for an additional 24 weeks. All patients underwent liver biopsy before and after the therapy period. Changes in cccDNA levels and liver histology were monitored between biopsies. Among the 11 patients who exhibited pre‐therapy hepatitis B e antigen (HBeAg), 8 (73%) showed evidence of HBeAg seroconversion by the end of the follow‐up period. Serum HBV DNA levels decreased by 5.2 and 3.3 log copies/ml (mean) by the end of the therapy and follow‐up periods, respectively. In addition, intrahepatic cccDNA decreased significantly to 1.4 log copies/µg (mean) by the end of the therapy period. Among the 11 patients who did not experience viral relapse, only 2 (18%) exhibited high levels of cccDNA (>4.5 log copies/µg) by the end of the treatment period. In contrast, all relapsed subjects exhibited significantly higher levels of cccDNA than subjects who did not relapse (P = 0.027). The combination regimen is a promising approach to treat chronic hepatitis B and may achieve significant reduction in serum HBV DNA and intrahepatic cccDNA. J. Med. Virol. 85: 987–995, 2013. © 2013 Wiley Periodicals, Inc.