HBV Genotype Research Articles

Clinical Features of Hepatitis B e-antigen Positive Chronic Hepatitis B Patients co-existing Precore and/or Basal Core Promoter Mutations

Aim: Characterize the clinical features of treatment-naive hepatitis B e-antigen (HBeAg)-positive chronic hepatitis B (CHB) patients harboring either precore (PC) or basal core promoter (BCP; PC/BCP) mutations. Methods: Alanine aminotransferase, HBsAg, HBV-DNA, HBV genotypes, PC/BCP mutations and liver stiffness measurement were assessed for 186 treatment-naive HBeAg-positive CHB patients. Results: PC/BCP mutations were found in 40.31% of the treatment-naive HBeAg-positive CHB patients. Patients with PC/BCP mutations showed significantly higher alanine aminotransferase and liver stiffness measurement levels. The serum HBsAg and HBV DNA levels in patients with BCP mutations were significantly lower than those in patients with wild-type virus. PC/BCP mutations were more prevalent in patients in immune clearance phase than those in immune tolerance phase. Conclusion: The emergences of PC/BCP mutations possibly indicate that HBeAg positive patients are experiencing the phase of immune clearance.

Using nucleotide sequencing to determine HBV genotypes in Kerman

Background: Hepatitis viruses are one of the serious medical problems and Hepatitis B is one of the chief transferable disease via blood and its products. Nowadays, 11 genotypes of hepatitis B have known over the world by the genome sequencing. Hepatitis B viruses have special geographical distribution. The clinical importance of hepatitis B viruses and its relation with the mutations has recognized. The purpose of this study was to check the presence and prevalence of Hepatitis B virus genotypes among the referrals attended to the medical diagnostic laboratories in Kerman province.Materials and Methods: In this cross-sectional study, twenty-one specimens were collected from blood samples available in the medical diagnostic laboratories of Kerman province during one year. After DNA extraction, PCR was carried on by specific primers, then they were sequenced. The obtained sequenced were compared with sequences in the NCBI gene bank and blasted for identification of their genotypes.Results: Seven samples from twenty one samples (33.3%) had D genotype, 13 samples from 21 (62%) had D3 subgenotype and 1 sample from 21 (4.7%) had D4 subgenotype.Conclusion: The prevalence of these genotypes in the Kermanian patients that recognized in this study can help to provide diagnostic kits for hepatitis B virus.Keywords: Hepatitis B virus, genotype, PCR, Sequencing, NCBI.

Prevalence and Genotypes of HBV and HCV and the risk Factors for co-infection with HIV-1 Among Patients attending Ngong Sub-County Hospital, Kenya

Prevalence and Genotypes of HBV and HCV and the risk Factors for co-infection with HIV-1 Among Patients attending Ngong Sub-County Hospital, Kenya

Host and Viral Genetic Variation in HBV-Related Hepatocellular Carcinoma.

Hepatocellular carcinoma (HCC) is the fifth most common cancer in men and the second leading cause of cancer deaths globally. The high prevalence of HCC is due in part to the high prevalence of chronic HBV infection and the high mortality rate is due to the lack of biomarkers for early detection and limited treatment options for late stage HCC. The observed individual variance in development of HCC is attributable to differences in HBV genotype and mutations, host predisposing germline genetic variations, the acquisition of tumor-specific somatic mutations, as well as environmental factors. HBV genotype C and mutations in the preS, basic core promoter (BCP) or HBx regions are associated with an increased risk of HCC. Genome-wide association studies have identified common polymorphisms in KIF1B, HLA-DQ, STAT4, and GRIK1 with altered risk of HBV-related HCC. HBV integration into growth control genes (such as TERT), pro-oncogenic genes, or tumor suppressor genes and the oncogenic activity of truncated HBx promote hepatocarcinogenesis. Somatic mutations in the TERT promoter and classic cancer signaling pathways, including Wnt (CTNNB1), cell cycle regulation (TP53), and epigenetic modification (ARID2 and MLL4) are frequently detected in hepatic tumor tissues. The identification of HBV and host variation associated with tumor initiation and progression has clinical utility for improving early diagnosis and prognosis; whereas the identification of somatic mutations driving tumorigenesis hold promise to inform precision treatment for HCC patients.

HBV epidemiology and genetic diversity in an area of high prevalence of hepatitis B in southern Brazil

BackgroundHepatitis B virus (HBV) infection is a major public health problem in Brazil. HBV endemicity is usually moderate to low according to geographic regions, and high prevalence of this virus has been reported in people of some specific Brazilian counties, including those with a strong influence of Italian colonization in southern Brazil. Analysis of HBV diversity and identification of the main risk factors to HBV infection are necessary to understand hepatitis B epidemiology in these high prevalence regions in southern Brazil. ObjectiveTo investigate epidemiological characteristics and HBV genotypes and subgenotypes circulating in a specific city with high HBV prevalence. MethodsA cross-sectional study was performed with 102 HBV chronically infected individuals, recruited in reference outpatient clinics for viral hepatitis in a city of high HBV prevalence (Bento Gonçalves) in Rio Grande do Sul state, Brazil between July and December 2010. Socio-demographic, clinical and behavior-related variables were collected in a structured questionnaire. HBV serological markers (HBsAg, anti-HBc), viral load, genotypes/subgenotypes and drug resistance were evaluated and comparatively analyzed among all patients. ResultsThe HBV infected subjects had a mean age of 44.9 (±12.2) years, with 86 patients (84.3%) reporting to have a family history of HBV infection, 51 (50.0%) to share personal objects, and were predominantly of Italian descendants (61; 64.9%). There was a predominance of genotype D (49/54; 90.7%), but genotype A was also detected (5/54; 9.3%). Subgenotypes D1 (1; 4.7%), D2 (3; 14.3%), and D3 (17; 81.0%) were identified. LAM-resistant mutation (rtM204I) and ADV-resistant mutations (rtA181V) were detected in only one patient each. ConclusionsThese results demonstrate a pivotal role of intrafamilial transmission for HBV spreading in this population. Furthermore, there is a high prevalence of HBV genotype D in this region.

Molecular epidemiology of hepatitis delta and hepatitis B viruses circulating in two major provinces (East and North-West) of Pakistan

ObjectivesHBV and HDV are major public health problems with millions affected globally and Pakistan accounts for a significant proportion of the global Hepatitis burden. This cross sectional study was designed to assess the general epidemiological and virological features of HBV and HDV in the Punjab and Khyber Pakhtunkhawa (KP) provinces of Pakistan. MethodsA total of 1890 HBV patients from March 2016 to May 2017 were recruited in the study and the presence of HDV was retrospectively evaluated in all participants. Most participants were young adults (from 21 to 30 years). Genotyping was based on PCR amplification using primers specific for HBV genotypes A-F, and HDV. 405 nucleotide fragments of HDV were sequenced. MEGA was used for phylogenetic analysis. ResultsOverall prevalence of HBV was 14.08% (266/1890). Higher prevalence was observed in males (66.85%) as compared to females (33.15%). Co-infection of HDV was found in 39 (14.66%) patients. HBV genotype-D was prevalent in dual infections followed by HDV/A (p < 0.05).While HDV genotype 1 was predominant in all HBV positive samples. Compared to Punjab, coinfection was higher in KP (14.3% versus15.2%; p < 0.05). ConclusionThe prevalence of HBV and HDV is high in Pakistan. The description of HBV and HDV genotypes circulating in East and North-West Pakistan can contribute to a better understanding of their relevance in regional epidemics. These infections are highly endemic in the KP, where their control is confounded by its vast territorial dimension with small, hard-to-reach municipalities and diverse ethnic populations.

Long-Term Follow-Up of Acute Hepatitis B: New Insights in Its Natural History and Implications for Antiviral Treatment.

Acute hepatitis B infection (AHB) is still a common viral acute hepatitis worldwide. As vaccination, antiviral treatment, and immigration are bound to affect the epidemiological landscape of HBV infections, and some of its aspects need to be investigated: (1) the circulation of vaccine escape mutants and of primary drug resistant strains; (2) the change in HBV genotype prevalence; and (3) the clinical implications of AHB and the probability of chronification. The serological, virological, and clinical parameters of 75 patients, acutely infected by HBV, were gathered for a retrospective study. Long-term follow up, either to complete seroconversion or for up to five years, was possible for 44 patients. Sequence analysis of the reverse transcriptase/HBsAg and precore regions was performed to investigate the molecular epidemiology and pathogenesis of recent infections by HBV. Genotype distribution in AHB in Italian patients was radically different from that of chronic infections, with a dramatic increase of extra-European genotypes (A1, F), suggesting that a proportion of AHBs are currently related to imported strains. None of the documented infections occurred in vaccinated individuals, while HBsAg variants (potentially vaccine escape variants) were rare and less prevalent than in chronic infections. No drug resistant strains were observed. Spontaneous viral clearance occurred in all but three cases. Time to viral clearance was inversely proportional to liver damage, but HBsAg titer on day 28 and, better still, HBsAg decay from day 0 to day 28 after admission, were the best predictors of chronification. They are, thus, potentially useful to guide antiviral treatment to prevent chronic evolution.

P2‐004: Molecular diversity of HBV and HDV amino‐acid sequences involved in HDV virion morphogenesis according to HBV and HDV genotypes: study of 1590 clinical strains

P2‐004: Molecular diversity of HBV and HDV amino‐acid sequences involved in HDV virion morphogenesis according to HBV and HDV genotypes: study of 1590 clinical strains

HBV molecular epidemiology and clinical condition of immigrants living in Italy.

We investigated 170 HBsAg-positive immigrants living in Italy for 1-7years to ascertain whether they may have become infected in the host country. Of 2032 adult immigrants interviewed, 1727 (85%) voluntarily adhered to a screening program for bloodborne or sexually transmitted infections. HBsAg was detected in 170 (9.8%) screened immigrants who completed the diagnostic, clinical and therapeutic process at the nearest clinic of infectious diseases. HBV molecular biology was performed applying a homemade technology. Phylogenetic signal of the datasets was obtained by a likelihood-mapping analysis using TreePuzzle. Of the 170 HBsAg-positive immigrants, 133 were inactive carriers, 29 had chronic hepatitis and 8 compensated cirrhosis. HBV genotype was identified in 109 of the 113 HBV-DNA-positive immigrants and HBV-genotype-E predominated (68.9%). Of these 109, 6 (5.5%) subjects showed an HBV genotype absent or extremely rare in their native country: HBV-genotype-E in three from Eastern Europe and in one from Sri Lanka, possibly acquired from other immigrants from sub-Saharan countries, HBV-genotype-D1 in one from Burkina Faso and one from Senegal, possibly acquired in Italy. The data suggest that immigrants may acquire HBV infection in Italy and, therefore, HBV vaccination programs should be extended to all immigrants living in Italy.

HBV genotyping based on key epitopes of PreS1 antigen and its correlation with genotyping by full-length PreS1 sequencing

Objective: The aim was to investigate the genotype distribution of two major epitopes of large surface protein (PreS1) of hepatitis B in Chinese patients and to explore the association between the genotypes of these two epitopes, and to determine whether PreS1 full-length genotype could be revealed according to the polypeptide sequence of key epitopes. Methods: HBV DNA was extracted from the serum of patients for PCR amplification. 278 samples amplified successfully were sequenced and compared with the known HBV sequences in Genbank to determine the two key epitopes of HBV PreS1 genotype (amino acid epitope 21-47 and 94-117, abbreviated as P21 and P94) and PreS1 full-length genotypes. The correlation among three genotyping approaches was analyzed by Cohen's kappa coefficient to verify the consistency between the key-epitope genotyping and the full-length preS1 genotyping. Results: 232 samples were successfully sequenced. The genotyping based on the kind of P21 epitope protein sequence, 201 cases for genotype C, 23 cases for genotype B and 8 cases for uncertain genotypes and genotyping based on the form of P94 epitope protein sequence, 199 cases for genotype C, 25 cases for genotype B and 8 cases for indeterminate genotypes. Lastly, the genotyping based on sequence of the full-length PreS1 sequence, 207 and 25 cases for genotype C and B. P21 or P94 epitope genotyping and PreS1 full length genotyping were highly consistent, respectively, 96.55% and 96.12%, and the two epitopes (P21and P94) genotyping have parallel consistency (93.10%). Conclusion: In this study, an innovatively genotyping method based on the amino acid sequence of key epitopes was proposed. The genotypes of HBV in china were mainly B and C genotypes, and the genotypes of key conserved epitopes of HBV PreS1 were highly consistent with the full-length genotyping ( > 96%). Moreover, genotyping with one or two key epitopes can be used in place of the full-length genotyping.

Sanguisorba officinalis extract, ziyuglycoside I, and II exhibit antiviral effects against hepatitis B virus

IntroductionSanguisorba officinalis (SO) has been commonly used to relieve dermatitis, burns, intestinal infections, hematemesis, and melena for a long time in China, Korea, and Japan. The aim of this study was to investigate the antiviral effects of 30% ethanol extract of SO (SOE) and its major compounds (ziyuglycoside I and II) against HBV in HepG2.2.15 cells and HepG2 cells transfected with HBV genotype C. MethodsThe antiviral activities of SOE were evaluated by ELISA and quantitative RT-PCR assay in in vitro models. In addition, its effect on HBsAg level was compared to entecavir (ETV), and its co-treatment with ETV was performed to observe combination efficacy. After HPLC analysis of ziyuglycoside I and II, the effects of those compounds on HBsAg levels were measured. ResultsSOE, ziyuglycoside I, and II significantly reduced HBsAg excretion and the pgRNA levels in HepG2.2.15 cells. In particular, SOE treatment of 100 μg/ml showed an inhibitory effect on HBsAg excretion similar to the levels found with ETV treatment of the same concentration. Additional use of 125 μg/ml of SOE in combination with 80 μM of ETV inhibited HBsAg production up to the level that is diminished by 320 μM ETV treatment in HepG2.2.15 cells. Among SOE, ziyuglycoside I, II, and ETV, ziyuglycoside II had the lowest inhibitory concentration of 50% against genotype D, and it was more potent against HBV genotype C than D in down-regulating HBsAg secretion. ConclusionTaken together, SOE noticeably inhibited replication and antigen secretion of HBV, which was at least in part attributed to ziyuglycoside II, thus presenting the possibility of developing therapeutic candidates for the treatment of HBV-related diseases.

Quantification of serum markers of hepatitis B (HBV) and Delta virus (HDV) infections in patients with chronic HDV infection.

The interplay between hepatitis B (HBV) and delta (HDV) viruses is complex and not always characterized during chronic HDV infection. We assessed the clinical usefulness of new quantitative assays for HBV and HDV serum markers in a retrospective cross-sectional study. Sera obtained from 122 HDV genotype 1 and HBV genotype D coinfected, anti-HIV-negative patients (71 males; median age 49.8 [21.7-66.9] years), recruited consecutively in two geographical areas (Italy 69 patients, Romania 53 patients) with different HBV and HDV epidemiology, were tested for HBsAg, HBV-DNA, HBcrAg, total anti-HBc, HDV-RNA, IgM and total anti-HDV using quantitative assays. Cirrhosis, which showed comparable prevalence in the two cohorts, was diagnosed in 97 of 122 (79.5%) patients. At multivariate analysis, cirrhosis was associated with lower total anti-HBc/IgM anti-HDV ratio (OR 0.990, 95% CI 0.981-0.999, P=.038), whereas disease activity was associated with higher total anti-HDV (OR 10.105, 95% CI 1.671-61.107, P=.012) and HDV-RNA levels (OR 2.366, 95% CI 1.456-3.844, P=.001). HDV-RNA serum levels showed a positive correlation with HBV-DNA (ρ=0.276, P=.005), HBsAg (ρ=0.404, P<.001) and HBcrAg (ρ=0.332, P<.001). The combined quantitative profiling of HBV and HDV serum markers identifies specific patterns associated with activity and stage of chronic hepatitis D (CHD). HDV pathogenicity depends on the underlying active HBV infection in spite of the inhibition of its replication. HDV-RNA, IgM anti-HDV, total anti-HDV, total anti-HBc, HBsAg and HBcrAg serum levels qualify for prospective studies to predict progressive CHD and identify candidates to antiviral therapy.

A Literature Review of Mathematical Models of Hepatitis B Virus Transmission Applied to Immunization Strategies From 1994 to 2015.

A mathematical model of the transmission dynamics of infectious disease is an important theoretical epidemiology method, which has been used to simulate the prevalence of hepatitis B and evaluate different immunization strategies. However, differences lie in the mathematical processes of modeling HBV transmission in published studies, not only in the model structure, but also in the estimation of certain parameters. This review reveals that the dynamics model of HBV transmission only simulates the spread of HBV in the population from the macroscopic point of view and highlights several main shortcomings in the model structure and parameter estimation. First, age-dependence is the most important characteristic in the transmission of HBV, but an age-structure model and related age-dependent parameters were not adopted in some of the compartmental models describing HBV transmission. In addition, the numerical estimation of the force of HBV infection did not give sufficient weight to the age and time factors and is not suitable using the incidence data. Lastly, the current mathematical models did not well reflect the details of the factors of HBV transmission, such as migration from high or intermediate HBV endemic areas to low endemic areas and the kind of HBV genotype. All of these shortcomings may lead to unreliable results. When the mathematical model closely reflects the fact of hepatitis B spread, the results of the model fit will provide valuable information for controlling the transmission of hepatitis B.

Concomitant Serum Presence of Hepatitis B Surface Antigen (HBsAg) and High Titers of Hepatitis B Surface Antibodies (Anti - HBsAb) in a Patient with Chronic Hepatitis B (HBV) Genotype D from Black Sea Coast Region: A Case Report

Introduction: Hepatitis B Virus genotypes influence chronic hepatitis B evolution and show geographic preferences. In Eastern Europe, HBV - D genotype seems to be dominant, still, without enough information regarding the prevalence of its four subtypes. In addition, treatment with Nucleos(t)ide Analogues (NUCs) has lower impact on HBV - D genotype clearance compared to others. This study aimed at presenting the case of a middle age female diagnosed, followed - up and treated for eight years with entecavir 0.5 mg/day, in whom an increase of anti - HBsAb titer was noted over the immunogenicity level and undetectable viremia, despite the continuous presence of HBsAg. This is an uncommon type of evolution, most patients with anti-HBsAb over 10 IU/mL show seroconversion in the “s” system in a few months. Case Presentation: The researchers used the COBAS TaqMan HBV Monitor Test (Roche Diagnostics, Branchburg, NJ) in order to measure serum HBV DNA level, then, the viral DNA was extracted from 200 μL of serum using QIAamp DNA blood mini kit (Qiagen, Germany). The amplification and sequencing of full DNA length was done by the Rolling Circle Amplification (RCA) technique. Hepatitis B Virus genotype was determined using the NCBI genotyping tool and phylogenetic analysis. Conclusions: The patient was proved to have D genotype. The DNA analyzes showed escape mutations in the “a” determinant within the S gene, represented by sQ129R, respectively, sI134T, reported by the literature in a small number of C genotype patients. The paradoxical serum profile of the current patient with positive HBsAg, increased titers of anti - HBsAb, and undetectable viral load could be a possible model of evolution for D genotype HBV chronic infected patients treated with NUCs, cohort genetic studies on patients with the same serum profile are required to confirm this pattern of evolution.

Evolution of HBV S-gene in the backdrop of HDV co-infection.

HBV-HDV co-infected people have a higher chance of developing cirrhosis, fulminant hepatitis, and hepatocellular carcinoma (HCC) compared to those infected only with HBV. The present study was conducted to investigate HBV genotypes and phylogeny among HBV mono-infected and HBV-HDV co-infected patients, as well as analyze mutations in the surface gene of HBV in mono-infected and co-infected patients. A total of 100 blood samples (50 co-infected with HBV and HDV, and 50 mono-infected with HBV only) were collected for this study. HBV DNA was extracted from patient sera and partial surface antigen gene was amplified from HBV genome using polymerase chain reaction. HBV S gene was sequenced from 49 mono-infected and 36 co-infected patients and analyzed to identify HBV genotypes and phylogenetic patterns. Subsequently, HBV S amino acid sequences were analyzed for mutational differences between sequences from mono- and co-infected patients. HBV genotype D was predominantly found in both mono-infected as well as co-infected patients. Phylogenetic analysis showed the divergence of HBV sequences, between mono- and co-infected patients, into two distinct clusters. HBV S gene mutation analysis revealed certain mutations in HBV-HDV co-infected subjects to be distinct from those found in mono-infected patients. This might indicate the evolution of HBV S gene under selection pressures generated from HDV coinfection.

Trends in hepatitis B virus resistance to nucleoside/nucleotide analogues in North China from 2009–2016: A retrospective study

Nucleos(t)ide analogues (NAs) are widely used in anti-hepatitis B virus (anti-HBV) therapy for effective inhibition of HBV replication. However, HBV resistance to NAs has emerged, resulting in virus reactivation and disease recurrence. Data on the current dynamics of HBV resistance are still rare in China. This study analysed 4491 plasma samples with HBV primary genotypic resistance mutations representative of the general HBV resistance situation in northern China from 2009-2016. We found that entecavir (ETV), representing 57.6% (12 713/22 060) of NA users in North China in 2016, has become the major NA for treating Chinese patients infected with HBV. Despite >50% of M204I/V±L180M among all HBV resistance cases annually and extensive exposure of patients to lamivudine (LAM), telbivudine (LdT) and adefovir dipivoxil (ADV), ETV resistance also showed a dramatically increased incidence, which rose to 17.1% in 2016. Moreover, A181T/V, ETV resistance mutations and multidrug resistance mutations were found more frequently in HBV genotype C compared with genotype B (21.2% vs. 8.5%, 12.4% vs. 7.9% and 5.9% vs. 3.0%, respectively), whereas M204I and N236T were more predominant in genotype B than genotype C (40.3% vs. 20.8% and 11.3% vs. 1.8%, respectively). In conclusion, we report the dynamic changes of HBV NA resistance mutation patterns and the current NA usage profile for anti-HBV treatment in North China over the past 8 years. These data provide valuable information on HBV NA resistance that is an important reference for clinicians to devise more effective treatment regimens for individual patients.

SAT-392 - Molecular diversity of HBV and HDV amino-acid sequences involved in HDV virion morphogenesis according to HBV and HDV genotypes: study of 1590 clinical strains

SAT-392 - Molecular diversity of HBV and HDV amino-acid sequences involved in HDV virion morphogenesis according to HBV and HDV genotypes: study of 1590 clinical strains

FRI-293 - Key mutational patterns in HBsAg C-terminus profoundly affect HBsAg levels in HBeAg-negative chronic HBV genotype D infection

FRI-293 - Key mutational patterns in HBsAg C-terminus profoundly affect HBsAg levels in HBeAg-negative chronic HBV genotype D infection

SAT-358 - Identification of HBV genotypes and mutations associated to antiviral resistance and vaccine escape in serum and oral fluid samples from chronic hepatitis B patients

SAT-358 - Identification of HBV genotypes and mutations associated to antiviral resistance and vaccine escape in serum and oral fluid samples from chronic hepatitis B patients

Comparison of liver function tests between different genotypes of HBV and HCV in non-responders of HBV-HCVCo-infected patients with healthy controls

Comparison of liver function tests between different genotypes of HBV and HCV in non-responders of HBV-HCVCo-infected patients with healthy controls