Evolution of precore/core promoter mutations in hepatitis B carriers with hepatitis B e antigen seroreversion

The evolution of precore stop codon mutation (A1896) and dinucleotide mutation (T1762/A1764) in the basic core promoter (BCP) of hepatitis B virus (HBV) genome during transient seroconversion and seroreversion of hepatitis B e antigen (HBeAg) remains unclarified. Five HBeAg-positive HBV carriers who experienced transient seroconversion followed by seroreversion of HBeAg (Group I, 3.3%) and 3 HBeAg-negative HBV carriers with documented reversion of HBeAg (Group II, 2.5%) in a prospective cohort of 272 patients with chronic hepatitis B were thus identified. The sequential changes at the precore nucleotide 1896 and BCP dinucleotide 1762/1764 were determined by polymerase chain reaction and direct sequencing. At enrollement, precore A1896 and BCP T1762/A1764 were noted in 4 (50%) and 1 (13%) of the eight patients. During a median follow-up period of 58 months (range: 31-76 months), 12 episodes of transient HBeAg seroconversion followed by seroreversion were encountered in Group I patients and 3 episodes of HBeAg seroreversion in Group II patients. Accompanying acute exacerbations were found in two-thirds of patients with either HBeAg seroconversion or seroreversion. Overall, precore nucleotide A1896 remained identical in 73% and 83% of the seroconversion and seroreversion events, respectively. BCP dinucleotide T1762/A1764 remained unchanged in 94% and 92% of the seroconversion and seroreversion events, respectively. At the end of follow-up, only one had both precore and BCP mutations. In conclusion, these data suggested that HBeAg seroreversion might be due to the lack of sustained precore and BCP mutations after HBeAg seroconversion. Although uncommon, HBeAg seroreversion can be associated with hepatitis exacerbation.     

Gender disparity in distribution of the major hydrophilic region variants of hepatitis B virus genotype C according to hepatitis B e antigen serostatus

Hepatitis B virus (HBV) e antigen (HBeAg) seroconversion during chronic HBV infection is known to play an important role in disease progression and patient response to antiviral agents. The aim of the present study was to analyze gender disparity in distribution of major hydrophilic region (MHR) variants according to HBeAg serostatus. Prevalence of MHR variants from 68 Korean patients with chronic hepatitis (31 HBeAg-positive and 37 HBeAg-negative) was examined in terms of HBeAg serostatus and sex by direct sequencing analysis of the MHR. Gender disparity was observed in the distribution of MHR variants according to HBeAg serostatus. In male patients, the prevalence of MHR variants was significantly higher in HBeAg negative patients than in HBeAg positive patients [58.8% (10/17 patients) vs. 14.3% (3/21 patients), P=0.004]. However, the same was not true in female patients [55.0% (11/20 patients) vs. 60.0% (6/10 patients), P=1.000)]. In addition, 2 mutation types (L110I and G145A) related to HBeAg serostatus were found. In conclusion, HBeAg seroconversion in male chronic patients infected with genotype C could lead to mutations of MHR, major target to host immune response, which might in turn contribute to HBV persistence and immune evasion.     

Viral genotypes and response to interferon in patients with acute prolonged hepatitis B virus infection of adulthood in Japan

Acute hepatitis B virus (HBV) infection was diagnosed in 57 adults admitted to Toranomon Hospital in Tokyo, Japan. Genotypes of HBV were determined by a serological method and compared to those in 1,077 patients with chronic hepatitis B. The distribution of genotypes were: genotype A (acute, 22.8% vs. chronic, 1.9%; P < 0.00001); B (14.0% vs. 9.4%); C (43.9% vs. 87.7%, P = 0.004); D (1.8% vs. 0.2%); F (1.8% vs. 0.2%); and unable to be typed (15.8% vs. 0.6%, P = 0.001). The infection persisted in seven (12%) of them. They included six (86%) of the seven patients who received prednisolone or glycyrrhizin during an acute phase of illness and one of the 41 (2%) who did not (P = 0.01). Interferon was given to the seven patients with acute prolonged HBV infection, and four of them responded by clearing hepatitis B e antigen (HBeAg) and surface antigen (HBsAg) from serum. Of the four responders, one was infected with HBV genotype B and three with genotype C. HBsAg persisted in the remaining three patients all of whom were infected with HBV genotype A, and HBeAg stayed positive in one of them. These results indicate that HBV genotype A prevails in Japanese patients with acute hepatitis B, and suggest a high efficacy of interferon in the adult patients with acute prolonged HBV infection, except in those infected with HBV genotype A.     

Detection of hepatitis B viral DNA by polymerase chain reaction in patients with hepatitis B surface antigen

Hepatitis B virus (HBV) DNA was assayed using the polymerase chain reaction in serum samples of 116 hepatitis B surface antigen (HBsAg) carriers, including 30 positive for hepatitis B e antigen (HBeAg) and 86 negative for HBeAg. In the HBeAg-positive group, all were positive for HBV DNA. In the HBeAg-negative group, 80.2% were positive for HBV DNA (80.0% in the healthy carrier group, 90.0% in the chronic active liver disease group, and 69.2% in patients with cirrhosis). This study indicated that every HBeAg-positive carrier as well as the majority of HBeAg-negative carriers were infectious and, in the latter group, that viral replication is most active in patients with chronic active liver disease.     

Virological outcomes in patients infected chronically with hepatitis B virus genotype A in comparison with genotypes B and C

In a single hospital in Tokyo, the 87 patients infected persistently with hepatitis B virus (HBV) genotype A, the 413 with B, and the 3,389 with C were compared for virological outcome. Hepatitis B surface antigen (HBsAg) was cleared from the serum in 12% (3/26), 2% (2/112), and 3% (23/826) of patients with genotypes A, B, and C, respectively, at 5 years of follow-up (P = 0.0395). Hepatitis B e antigen (HBeAg) was cleared from serum more frequently in patients with genotype B than those with A or C (78% [32/41] vs. 58% [11/19] or 45% [251/562], P = 0.00001) at 5 years. Of the 45 individuals infected with genotype A and followed for 3 years or longer, HBeAg was more frequent (16% [3/19] vs. 73% [19/26], P = 0.0002) and levels of HBV DNA higher (median <2.6 [range: <2.6-5.6] vs. >7.6 [<2.6->7.6] log copies/ml, P = 0.001) in the 26 patients with biopsy-proven chronic hepatitis than the 19 asymptomatic carriers. Among the 26 hepatitis patients infected with HBV genotype A, decreases in HBV DNA were less frequent (20% [1/5] vs. 93% [13/14] or 86% [6/7], P = 0.0095) and increases in serum levels of hyaluronic acid > or =10 ng/ml commoner (80% [4/5] vs. 14% [2/14] or 14% [1/7], P = 0.017) in the patients who kept HBeAg than in those who seroconverted or who remained HBeAg-negative. In conclusion, patients persistently infected with HBV genotype A fare better than those with genotype B or C. However, high levels of HBV DNA continue in those in whom HBeAg persists along with fibrosis in the liver.     

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.     

Hepatitis B virus genotypes and spontaneous hepatitis B e antigen seroconversion in Taiwanese hepatitis B carriers

Hepatitis B virus (HBV) is classified into eight genotypes (A-H), and genotype C is associated with more aggressive liver disease compared to genotype B. However, the mechanisms responsible for the clinical differences remain unclear. To test whether genotype C patients had with lower rates of spontaneous hepatitis B ge antigen (HBeAg) seroconversion than genotype B patients, stored serum samples from 146 Taiwanese adult HBeAg-positive hepatitis B carriers followed-up for a mean of 52 months (range, 12-120 months) were tested for HBV genotype by a molecular method. Genotype C patients were significantly older than genotype B patients (mean age, 37 +/- 12 vs. 29 +/- 10 years, P < 0.001). During the follow-up period, genotype C patients had a significantly lower rate of spontaneous HBeAg seroconversion than genotype B patients (27 vs. 47%, P < 0.025). Spontaneous HBeAg seroconversion occurred one decade later in genotype C patients compared with genotype B patients. Multivariate analyses identified age < or =35 years (odds ratio: 2.08; 95% confidence interval [CI], 1.07-4.0; P < 0.05), high baseline serum alanine aminotransferase level (odds ratio: 2.34; 95%CI, 1.39-4.09; P < 0.005), and HBV genotype B (odds ratio: 1.94; 95%CI, 1.03-3.63; P < 0.05) as independent factors associated with spontaneous HBeAg seroconversion. In conclusion, genotype C patients, compared to genotype B patients, have a delayed HBeAg seroconversion in the immune clearance phase of chronic HBV infection, which may contribute to a more progressive liver disease and more refractory to antiviral therapy.     

Comparison of genotypes C and D of the hepatitis B virus in Japan: a clinical and molecular biological study

The genotype-related differences between genotype C and genotype D of the hepatitis B virus (HBV) remain unknown. The relationship was studied between the HBV genotypes and their clinical features, paying special attention to genotypes C and D. Serum samples from 413 HBV carriers were genotyped using an enzyme immunoassay (EIA) and by restriction fragment length polymorphism. The nucleotide sequences at the basic core promoter (BCP) and precore (PreC) regions were analysed by direct sequencing. The full genome sequences of three HBV genotype D cases were also examined. Almost all carriers with HBV genotype D were asymptomatic carriers (84.2%). Genotype D was not found in patients with liver cirrhosis and hepatocellular carcinoma. In contrast, carriers with genotype C had mainly chronic liver disease (63.2%; P<0.001). The ratio of hepatitis B e antigen (HBeAg)/anti-HBe was significantly higher in genotype C than in genotype D in the young age-matched group (P<0.01). The mutation at BCP (T1762, A1764) was significantly lower in genotype D than in genotype C among HBeAg-negative patients (P<0.05). The HBV full-genome sequences are very similar to certain HBV genotype D sequences from Europe. In conclusion, genotype C was associated with chronic liver disease, whereas genotype D was related to asymptomatic carriers with earlier HBeAg seroconversion. Thus, the outcome of chronic HBV infection may be different in persons infected with HBV genotypes C and D.     

HBeAg阴性与阳性慢性乙肝患者的临床对比分析

目的 探讨HBeAg阳性和HBeAg阴性慢性乙肝患者的临床特征异同.方法 随机选取慢性乙肝患者354例,其中HBeAg阳性组124例,HBeAg阴性组230例,对两组的人口学、生化学、病毒学及诊断分型进行分析比较.结果 ①与HBeAg阳性组比较,HBeAg阴性组患者年龄较大(P=0.000),中度和重度慢性肝炎比例较低(P:0.007和0.014),重型肝炎发生率较高(P=0.008).②HBeAg阴性组的ALT、ALB、PTA及HBV DNA载量对数值低于HBeAg阳性组(P=0.035,0.002,0.000和0.000),但TBil水平高于HBeAg阳性组(P=0.003);两组AST水平差异无统计学意义(P=0.222).③HSeAs阴性组在高病毒载量组(HBV DNA105拷贝/m1)的比例低于HBeAg阳性组(37.4%VS55.6%,P=0.001).结论 HBeAg阴性患者与HBeAg阳性患者相比存在年龄偏大和HBV DNA水平较低等特征,HBeAg阴性乙肝患者的病情有时反而可能较重.     

HBeAg阴性与阳性慢性乙肝患者的临床对比分析

目的 探讨HBeAg阳性和HBeAg阴性慢性乙肝患者的临床特征异同.方法 随机选取慢性乙肝患者354例,其中HBeAg阳性组124例,HBeAg阴性组230例,对两组的人口学、生化学、病毒学及诊断分型进行分析比较.结果 ①与HBeAg阳性组比较,HBeAg阴性组患者年龄较大(P=0.000),中度和重度慢性肝炎比例较低(P:0.007和0.014),重型肝炎发生率较高(P=0.008).②HBeAg阴性组的ALT、ALB、PTA及HBV DNA载量对数值低于HBeAg阳性组(P=0.035,0.002,0.000和0.000),但TBil水平高于HBeAg阳性组(P=0.003);两组AST水平差异无统计学意义(P=0.222).③HSeAs阴性组在高病毒载量组(HBV DNA105拷贝/m1)的比例低于HBeAg阳性组(37.4%VS55.6%,P=0.001).结论 HBeAg阴性患者与HBeAg阳性患者相比存在年龄偏大和HBV DNA水平较低等特征,HBeAg阴性乙肝患者的病情有时反而可能较重.     

HBeAg阴性与阳性慢性乙肝患者的临床对比分析

目的 探讨HBeAg阳性和HBeAg阴性慢性乙肝患者的临床特征异同.方法 随机选取慢性乙肝患者354例,其中HBeAg阳性组124例,HBeAg阴性组230例,对两组的人口学、生化学、病毒学及诊断分型进行分析比较.结果 ①与HBeAg阳性组比较,HBeAg阴性组患者年龄较大(P=0.000),中度和重度慢性肝炎比例较低(P:0.007和0.014),重型肝炎发生率较高(P=0.008).②HBeAg阴性组的ALT、ALB、PTA及HBV DNA载量对数值低于HBeAg阳性组(P=0.035,0.002,0.000和0.000),但TBil水平高于HBeAg阳性组(P=0.003);两组AST水平差异无统计学意义(P=0.222).③HSeAs阴性组在高病毒载量组(HBV DNA105拷贝/m1)的比例低于HBeAg阳性组(37.4%VS55.6%,P=0.001).结论 HBeAg阴性患者与HBeAg阳性患者相比存在年龄偏大和HBV DNA水平较低等特征,HBeAg阴性乙肝患者的病情有时反而可能较重.     

HBeAg阴性与阳性慢性乙肝患者的临床对比分析

目的 探讨HBeAg阳性和HBeAg阴性慢性乙肝患者的临床特征异同.方法 随机选取慢性乙肝患者354例,其中HBeAg阳性组124例,HBeAg阴性组230例,对两组的人口学、生化学、病毒学及诊断分型进行分析比较.结果 ①与HBeAg阳性组比较,HBeAg阴性组患者年龄较大(P=0.000),中度和重度慢性肝炎比例较低(P:0.007和0.014),重型肝炎发生率较高(P=0.008).②HBeAg阴性组的ALT、ALB、PTA及HBV DNA载量对数值低于HBeAg阳性组(P=0.035,0.002,0.000和0.000),但TBil水平高于HBeAg阳性组(P=0.003);两组AST水平差异无统计学意义(P=0.222).③HSeAs阴性组在高病毒载量组(HBV DNA105拷贝/m1)的比例低于HBeAg阳性组(37.4%VS55.6%,P=0.001).结论 HBeAg阴性患者与HBeAg阳性患者相比存在年龄偏大和HBV DNA水平较低等特征,HBeAg阴性乙肝患者的病情有时反而可能较重.     

HBeAg阴性与阳性慢性乙肝患者的临床对比分析

目的 探讨HBeAg阳性和HBeAg阴性慢性乙肝患者的临床特征异同.方法 随机选取慢性乙肝患者354例,其中HBeAg阳性组124例,HBeAg阴性组230例,对两组的人口学、生化学、病毒学及诊断分型进行分析比较.结果 ①与HBeAg阳性组比较,HBeAg阴性组患者年龄较大(P=0.000),中度和重度慢性肝炎比例较低(P:0.007和0.014),重型肝炎发生率较高(P=0.008).②HBeAg阴性组的ALT、ALB、PTA及HBV DNA载量对数值低于HBeAg阳性组(P=0.035,0.002,0.000和0.000),但TBil水平高于HBeAg阳性组(P=0.003);两组AST水平差异无统计学意义(P=0.222).③HSeAs阴性组在高病毒载量组(HBV DNA105拷贝/m1)的比例低于HBeAg阳性组(37.4%VS55.6%,P=0.001).结论 HBeAg阴性患者与HBeAg阳性患者相比存在年龄偏大和HBV DNA水平较低等特征,HBeAg阴性乙肝患者的病情有时反而可能较重.     

HBeAg阴性与阳性慢性乙肝患者的临床对比分析

目的 探讨HBeAg阳性和HBeAg阴性慢性乙肝患者的临床特征异同.方法 随机选取慢性乙肝患者354例,其中HBeAg阳性组124例,HBeAg阴性组230例,对两组的人口学、生化学、病毒学及诊断分型进行分析比较.结果 ①与HBeAg阳性组比较,HBeAg阴性组患者年龄较大(P=0.000),中度和重度慢性肝炎比例较低(P:0.007和0.014),重型肝炎发生率较高(P=0.008).②HBeAg阴性组的ALT、ALB、PTA及HBV DNA载量对数值低于HBeAg阳性组(P=0.035,0.002,0.000和0.000),但TBil水平高于HBeAg阳性组(P=0.003);两组AST水平差异无统计学意义(P=0.222).③HSeAs阴性组在高病毒载量组(HBV DNA105拷贝/m1)的比例低于HBeAg阳性组(37.4%VS55.6%,P=0.001).结论 HBeAg阴性患者与HBeAg阳性患者相比存在年龄偏大和HBV DNA水平较低等特征,HBeAg阴性乙肝患者的病情有时反而可能较重.     

HBeAg阴性与阳性慢性乙肝患者的临床对比分析

目的 探讨HBeAg阳性和HBeAg阴性慢性乙肝患者的临床特征异同.方法 随机选取慢性乙肝患者354例,其中HBeAg阳性组124例,HBeAg阴性组230例,对两组的人口学、生化学、病毒学及诊断分型进行分析比较.结果 ①与HBeAg阳性组比较,HBeAg阴性组患者年龄较大(P=0.000),中度和重度慢性肝炎比例较低(P:0.007和0.014),重型肝炎发生率较高(P=0.008).②HBeAg阴性组的ALT、ALB、PTA及HBV DNA载量对数值低于HBeAg阳性组(P=0.035,0.002,0.000和0.000),但TBil水平高于HBeAg阳性组(P=0.003);两组AST水平差异无统计学意义(P=0.222).③HSeAs阴性组在高病毒载量组(HBV DNA105拷贝/m1)的比例低于HBeAg阳性组(37.4%VS55.6%,P=0.001).结论 HBeAg阴性患者与HBeAg阳性患者相比存在年龄偏大和HBV DNA水平较低等特征,HBeAg阴性乙肝患者的病情有时反而可能较重.     

HBeAg阴性与阳性慢性乙肝患者的临床对比分析

目的 探讨HBeAg阳性和HBeAg阴性慢性乙肝患者的临床特征异同.方法 随机选取慢性乙肝患者354例,其中HBeAg阳性组124例,HBeAg阴性组230例,对两组的人口学、生化学、病毒学及诊断分型进行分析比较.结果 ①与HBeAg阳性组比较,HBeAg阴性组患者年龄较大(P=0.000),中度和重度慢性肝炎比例较低(P:0.007和0.014),重型肝炎发生率较高(P=0.008).②HBeAg阴性组的ALT、ALB、PTA及HBV DNA载量对数值低于HBeAg阳性组(P=0.035,0.002,0.000和0.000),但TBil水平高于HBeAg阳性组(P=0.003);两组AST水平差异无统计学意义(P=0.222).③HSeAs阴性组在高病毒载量组(HBV DNA105拷贝/m1)的比例低于HBeAg阳性组(37.4%VS55.6%,P=0.001).结论 HBeAg阴性患者与HBeAg阳性患者相比存在年龄偏大和HBV DNA水平较低等特征,HBeAg阴性乙肝患者的病情有时反而可能较重.     

HBeAg阴性与阳性慢性乙肝患者的临床对比分析

目的 探讨HBeAg阳性和HBeAg阴性慢性乙肝患者的临床特征异同.方法 随机选取慢性乙肝患者354例,其中HBeAg阳性组124例,HBeAg阴性组230例,对两组的人口学、生化学、病毒学及诊断分型进行分析比较.结果 ①与HBeAg阳性组比较,HBeAg阴性组患者年龄较大(P=0.000),中度和重度慢性肝炎比例较低(P:0.007和0.014),重型肝炎发生率较高(P=0.008).②HBeAg阴性组的ALT、ALB、PTA及HBV DNA载量对数值低于HBeAg阳性组(P=0.035,0.002,0.000和0.000),但TBil水平高于HBeAg阳性组(P=0.003);两组AST水平差异无统计学意义(P=0.222).③HSeAs阴性组在高病毒载量组(HBV DNA105拷贝/m1)的比例低于HBeAg阳性组(37.4%VS55.6%,P=0.001).结论 HBeAg阴性患者与HBeAg阳性患者相比存在年龄偏大和HBV DNA水平较低等特征,HBeAg阴性乙肝患者的病情有时反而可能较重.     

HBeAg阴性与阳性慢性乙肝患者的临床对比分析

目的 探讨HBeAg阳性和HBeAg阴性慢性乙肝患者的临床特征异同.方法 随机选取慢性乙肝患者354例,其中HBeAg阳性组124例,HBeAg阴性组230例,对两组的人口学、生化学、病毒学及诊断分型进行分析比较.结果 ①与HBeAg阳性组比较,HBeAg阴性组患者年龄较大(P=0.000),中度和重度慢性肝炎比例较低(P:0.007和0.014),重型肝炎发生率较高(P=0.008).②HBeAg阴性组的ALT、ALB、PTA及HBV DNA载量对数值低于HBeAg阳性组(P=0.035,0.002,0.000和0.000),但TBil水平高于HBeAg阳性组(P=0.003);两组AST水平差异无统计学意义(P=0.222).③HSeAs阴性组在高病毒载量组(HBV DNA105拷贝/m1)的比例低于HBeAg阳性组(37.4%VS55.6%,P=0.001).结论 HBeAg阴性患者与HBeAg阳性患者相比存在年龄偏大和HBV DNA水平较低等特征,HBeAg阴性乙肝患者的病情有时反而可能较重.     

Response to long-term lamivudine treatment in patients infected with hepatitis B virus genotypes A, B, and C

Response to lamivudine treatment longer than 1 year was compared in 15 patients persistently infected with hepatitis B virus (HBV) genotype A, 38 with genotype B, and 449 with genotype C. Patients with genotype A were younger (median age 37 [range 24-49] vs. 47 [24-67] or 44 [18-73], P = 0.015), possessed hepatitis B e antigen (HBeAg) more frequently (73% vs. 21% or 56%, P < 0.001) and HBV DNA in higher levels (8.6 [6.1-8.7] vs. 6.5 [<3.7-8.7] or 6.5 [<3.7-8.7] log genome equivalents (LGE)/ml, P = 0.024) than those with genotype B or C. During lamivudine, YMDD mutants (89% vs. 53% or 42%, P = 0.0001) and breakthrough hepatitis developed more often (47% vs. 21% or 29%, P = 0.023) in patients with genotype A than B or C. YMDD mutants elicited more frequently in patients with genotype A than B or C who were positive (82% [9/11] vs. 25% [2/8] or 48% [117/245], P = 0.037) or negative for HBeAg (75% [3/4] vs. 30% [9/30] or 33% [68/204], P = 0.003). HBeAg (hazard ratio 2.1 [95% confidence interval 1.53-2.92], P < 0.001) and genotype A (2.78 [1.08-7.12], P = 0.034) enhanced the emergence of YMDD mutants by the Cox proportional hazard model. The risk for breakthrough hepatitis was increased by the baseline alanine aminotransferase level <500 IU/L (2.56 [1.82-5.50], P = 0.018), HBeAg (2.11 [1.40-3.16], P < 0.001), cirrhosis (1.92 [1.24-2.97], P = 0.004) and HBV DNA > or =8.0 LGE/ml (1.57 [1.04-2.36], P = 0.03); it was influenced by genotypes only in patients with HBeAg. In conclusion, HBV genotypes help in predicting response to long-term lamivudine treatment and development of YMDD mutants in patients with chronic hepatitis B.     

Evolutionary perspective on hepatitis B virus with an expanded sampling strategy

To investigate the role hepatitis B e antigen (HBeAg) plays in the evolution of hepatitis B virus (HBV), we sequenced the basic core promoter (BCP) and precore (preC) regions of 348 clones total from ten HBV Chinese patients. Eleven mutations were more frequent in HBeAg-negative patients than in HBeAg-positive patients. Further, the sequencing of dozens of variants was found to be necessary to obtain mutation profiles. Phylogenetic and median-joining network analyses suggested that variants from each patient had a single common ancestor (monophyly). Higher haplotype and nucleotide diversities were identified in HBeAg-negative patients. Analysis of dN/dS suggested that viruses experiencing a stronger immune response had lower haplotype diversity. Because HBeAg seroconversion was associated with viral diversity it served as an indicator of HBV evolution. Significantly, this study indicated a larger sampling of variants from each patient was required to understand effectively the properties of HBV.