HBV Drug Resistance Research Articles

Analysis of characteristics of drug resistance gene mutation in HBV RT region of hepatitis B infected patients

Objective: This article investigated the clinical characteristics and distribution of drug resistance mutation sites in HBV RT region of hepatitis B infected patients. Methods: Retrospective analysis was made on 1 948 patients with HBV infection, who had been tested for NAs resistance mutation and had a medical history of NAs in the Laboratory Department of the Fifth Medical Center of the PLA General Hospital from January 2020 to December 2021. Basic clinical information and drug resistance related mutation information were recorded. Meanwhile, the serological index data of hepatitis B were collected. Drug resistance gene mutant group and non-mutated group were grouped according to whether the drug resistance genes had a mutation in HBV RT region, and the clinical characteristics and genotype distribution of the two groups were statistically analyzed. The pattern of drug resistance gene mutation, number of mutation sites, drug resistance type and mutation of NAs resistance-related sites were analyzed in 917 patients with drug resistance gene mutation in HBV RT region. χ2 Inspection was used for counting data. Meanwhile, two independent samples t-test and Wilcoxon rank sum test were used for measurement data. Results: Among the 1 948 patients with chronic HBV infection, 917 patients had drug resistance gene mutation in RT region (47.07%). The proportion of patients with acute hepatitis B and CHB in HBV RT resistance gene mutant group was lower than that in the non-mutated group, while the proportion of patients with HBV-related cirrhosis was higher than that in the non-mutated group, these differences were statistically significant. Compared with the non-mutated group in HBV RT region, the age, the positive rates of HBeAg and HBV DNA, and HBV DNA load of these patients were increased in drug resistance gene mutant group, these differences were statistically significant. Genotypes of patients in both groups were dominated by C, followed by B and D. The proportion of patients with genotype C in HBV RT drug resistance gene mutant group was higher than that of non-mutated group, the difference was statistically significant. There were 53 gene mutation patterns in 917 patients with drug resistance gene mutation in HBV RT region, and the main pattern was rtL180M+rtM204V+rtS202G (9.70%). The mutation sites were dominated by 3 (20.74%). There were 5 types of drug resistance, LAM+Ldt (21.25%) was the most. Among the 18 sites that were clearly associated with LAM, ADV, ETV and Ldt resistance in the HBV RT region, 14 sites were mutated, and the most common mutation sites were rtL180M, rtM204V, rtM204 and rtS202G. what's more, the proportion of patients with NAs drug resistance was LAM>Ldt>ETV>ADV. Conclusion: In order to prevent adverse consequences of this study such as disease recurrence or disease progression caused by HBV drug resistance, HBV infected patients, who have long-term use of NAs antiviral therapy, should monitor the level of HBV DNA and drug resistance genes in HBV RT region in order to optimize the treatment plan in time or guide individualized treatment.

Resistant mutations within the hepatitis B virus reverse transcriptase sequence in treatment failure patients with chronic HBV infection in Vietnam

ObjectivesWe conducted this study to describe whether mutations in the gene coding for the enzyme reverse transcriptase (RT) were related to drugs used in the treatment of hepatitis B in Vietnam. MethodsPatients receiving antiretroviral therapy with evidence of treatment failure were included in the study. The RT fragment was cloned using the polymerase chain reaction technique after being extracted from patients’ blood samples. The nucleotide sequences were analysed using Sanger method. The HBV drug resistance database contains mutations associated to resistance to existing HBV therapies. Medical records were accessed to collect information on patient parameters, such as treatment, viral load, biochemistry, and blood count. ResultsResistance mutations to lamivudine, telbivudine, and entecavir were found in the highest proportion (75–91.7%) of HBV samples from patients who had failed antiretroviral therapy. Only 20.8% of HBV strains had mutations exhibiting adefovir resistance, while none had mutations conferring tenofovir resistance. M204I/V, L180M, and L80I are frequent variants linked with resistance to lamivudine, telbivudine, and entecavir. In contrast, the A181L/T/V mutation was detected predominantly in tenofovir-resistant HBV strains. Following the drug resistance mutation test, patients achieved the greatest virological response after 24 weeks of therapy with tenofovir and entecavir at a daily dose of one tablet. ConclusionLamivudine, telbivudine, and entecavir were all highly resistant to the RT enzyme modifications in 24 treatment failure patients, with M204I/V, L180M, and L80I being the most prevalent mutations. Tenofovir resistance mutations have not been found in Vietnam.

Prevalence and characteristics of hepatitis B and D virus infections among HIV-positive individuals in Southwestern Nigeria

BackgroundCoinfections of HIV-positive individuals with Hepatitis B and D virus (HBV and HDV) are common and can be associated with rapid liver damage. Several antiretroviral drugs for HIV exhibit anti-HBV effect; however, the selection of HBV drug resistance mutations (DRMs) in individuals under HIV antiretroviral therapy (ART) has been reported but rarely in Nigeria. In this study the HBV/HDV prevalence and HBV DRMs in HIV-positive individuals in Southwestern Nigeria were assessed.MethodsPlasma samples collected from 310 HIV-positive individuals including 295 ART-experienced and 15 ART-naïve persons attending the HIV clinic in three south-western states of Nigeria between June 2017 and August 2017 were analysed by ELISA for HBsAg and anti-HDV. The presence of HDV RNA and HBV DNA was analysed by (RT)-PCR followed by sequencing and phylogenetic analyses for genotyping. The HBV reverse transcription (RT) region was amplified and sequenced for the analysis of drug resistance mutations.ResultsOverall, 16.1% (n = 50/310) of the HIV-positive individuals were positive for HBsAg, most of which were ART-experienced (94.0%; n = 47/50). From the 50 HBsAg-positive samples, 72.0% (n = 36/50) were positive for HBV DNA and 16.0% (n = 8/50) had detectable HDV RNA while 5.6% (n = 2/36) of the HBV-DNA positive samples had anti-HDV total antibodies. Sequences were available for 31/36 of the HBV DNA-positive and 3/8 HDV RNA-positive samples. HBV DNA-positive samples were characterised as HBV genotype E infections exclusively, while HDV genotype 1 was detected in the HDV RNA-positive samples. HBV DRMs V173L, L180M, S202I and M204V/I, which are associated with lamivudine resistance, were detected in 32.2% (n = 10/31) of the HBV DNA-positive samples. Most of these mutations (90.0%; n = 9/10) were present in the ART-experienced cohort.ConclusionsThis study indicates that HBV/HDV coinfections are common in HIV-positive individuals under ART in Nigeria. Furthermore, a high proportion of HBV DRMs which potentially compromise future treatment options were detected, underscoring the need for HBV screening prior to starting ART. Further studies should be performed to monitor a possible increase in the spread of HDV among populations at risk of HIV and HBV infections.

Prevalence and genetic variability of occult hepatitis B virus in a human immunodeficiency virus positive patient cohort in Gondar, Ethiopia.

Occult hepatitis B (OHB) is a major concern in HIV infected patients as it associates with a high risk of HBV reactivation and disease progression. However, data on the prevalence of OHB among HIV positive patients in Ethiopia is lacking. This study aims to determine the prevalence of OHB in HBV/HIV co-infected patients from Gondar, Ethiopia. A total of 308 consented HIV positive patients were recruited from the University of Gondar Teaching Hospital, Ethiopia. Clinical and demographic data of the participants were recorded. Plasma was tested for HBsAg and anti-HBc using commercial assays (Abbott Architect). In HBsAg negative anti-HBc positive patient samples, total DNA was isolated and amplified using nested PCR with primers specific to HBV polymerase, surface and pre-core/core regions, followed by Sanger sequencing and HBV mutational analysis using MEGA 7.0. Of the total study subjects, 62.7% were female, median age 38.4 years, interquartile range (IQR): 18-68, and 208 (67.5%) had lifestyle risk factors for HBV acquisition. Two hundred and ninety-one study subjects were HIV+/HBsAg-, out of which 115 (39.5%) were positive for anti-HBc. Occult hepatitis B was detected in 19.1% (22/115) of anti-HBc positive HIV patients. HBV genotype D was the predominant genotype (81%) among OHB positive patients. Mutations associated with HBV drug resistance, HBV reactivation, and HCC risk were detected in 23% (5/22), 14% (3/22) and 45.5% (10/22) of patients, respectively. This study found a high rate of occult hepatitis B in HIV patients. Further, high rates of mutations associated with HBV reactivation, drug resistance, and HCC risk were detected in these patients. These data highlighted the need for integrating OHB screening for proper management of liver diseases in HIV patients.

Tenofovir plus entecavir combination therapy for chronic hepatitis B with nucleos(t)ide analogue failure

Objectives: Currently, both entecavir (ETV) and tenofovir (TDF) are recommended first-line therapy for chronic hepatitis B treatment due to good tolerance, lower side-effect profile and high genetic barrier. However, mutations that may develop in the polymerase gene during treatment may result treatment failure. In this study, we aimed to evaluate the efficacy and virologic response of ETV plus TDF combination therapy in chronic hepatitis B patients with suboptimal response to nucleos(t)ide analogues (NAs) monotherapy. Methods: A total of 813 patients who were followed-up with the diagnosis of chronic hepatitis B and who were treated with TDF or ETV monotherapy were screened. Patients who had a partial or non response to monotherapy during at least 12 months and who was the presence of serum HBV-DNA levels ≥ 2000 IU/mL at the time of initiation of the ETV plus TDF combination therapy were included. Results: Ten (1.2%) patients (9 TDF, 1 ETV) were identified to have had partial response (50%) or breaktrough (40%) or virologic rebound (10%) to monotherapy. The median age was 36.8 years (range, 22-55 years), and 5/10 (50%) patients were male. Of 10 patients, nine of achieved undetectable HBV-DNA (< 15 IU/mL) levels (50% of in 6 months and 90% of in 18 months) with combination therapy. One patient showed no response. Conclusions: Our results suggest that combination therapy is superior to the antiviral change in treatment failure with NAs. In addition, it is important to conduct HBV drug resistance analyzes to prevent false drug change in treatment.

Characterization of Hepatitis B Virus and Human Immunodeficiency Virus among HIV/HBV Co-Infected Patients from the Republic of Guinea

Aim. Molecular genetic characterization of hepatitis B virus and human immunodeficiency virus in patients with HIV / HBV co-infection living in the Republic of Guinea. Materials and methods. 2168 blood serum samples obtained from the Republic of Guinea residents – blood donors and conditionally healthy people, without suspicion of Ebola virus disease, UK RUSAL employees and their families, as part of their routine medical examination. The presence of serological and molecular biological markers of HIV and HBV was examined. When HIV/HBV co-infection was detected, the nucleotide sequences of the complete HBV genomes and the HIV pol gene fragment were sequenced. Results and discussion. HIVserological markers were detected in 239 people (11.02 %). HIV RNA was detected in 31 people, which accounted for 12.9 % of patients in the seropositive group (1.43 % of the total group). HBV serological markers among HIV RNAs-positive individuals were detected in 29.03 % of patients, including 16.12 % HBsAg and 12.9 % anti-HBcore IgG. HBV DNA was detected in all HBsAg-positive and in two anti-HBcore IgG-positive patients, as well as in 12 people negative for all HBV serological markers analyzed in the work. Thus, HBV DNA was found in 61.29 % of HIV RNA-positive individuals. Based on the pol gene fragment nucleotide sequences analysis of 19 HIV samples, it was shown that the HIV circulating recombinant form CRF02_AG prevails in the examined group (52.63 %) compared with HIV A1 (42.1 %), one sample was an independent recombinant of genotypes A1 and G. HBV phylogenetic analysis of the studied samples showed that genotype E prevails – 47.36 %, compared with HBV D1 – 21.05 %, D2 – 15.78 %, D3 – 10.52 % and A2 – 5.26 %. HIV and HBV samples have been detected that carry drug resistance mutations despite the antiretroviral therapy absence. HIV and HBV drug resistance mutations identification in ART-naive patients emphasizes the need for HIV surveillance programs as well as routine testing for HBV and HIV and HBV drug resistance before starting antiretroviral therapy in the clinical management of patients in the country.

Identification and molecular-genetic characteristics of the hepatitis B virus among HIV-infected patients in Arkhangelsk.

To estimate the prevalence and characterize the hepatitis B virus among HIV-infected patients with virological failure of antiretroviral therapy in Arkhangelsk. HBV markers determinations (HBsAg, anti-HBs IgG, antiHBcor IgG, DNA HBV) were performed in isolates from blood plasma samples 64 HIV-infected patients with virological failure of antiretroviral therapy (viral load >50 IU / ml after 6 months of antiretroviral therapy or an increase in viral load after primary suppression of viral replication). For the detection of the hepatitis B virus, nucleic acids were isolated using the commercial kit «AmplePrime Ribo-prep». The virus presence analysis was performing by the polymerase chain reaction (PCR) method with hybridization-fluorescence detection in "real time" using the commercial set of «AmpliSens® HBV-FL». In the future, we used the method developed by the Saint-Petersburg Pasteur Institute, which allows detecting HBV in biological material with a low viral load. HBsAg-negative (occult) HBV was detect in 28 (43.8%) HIVinfected patients. Only HBV genotype D was detected, and the HBV subgenotype D1 prevailed (39.3%) compared with the HBV subgenotype D2 (32.1%) and D3 (28.6%). Serological markers in 42.8% of patients with HBV DNA were founding. Two HBV isolates with drug resistance mutations in the polymerase gene, leaded to amino acid substitutions (L180M, M204V) associated with the resistance development to lamivudine, entecavir, telbivudine and tenofovir were identifying. The occult (HBsAg-negative) HBV high prevalence among HIV-infected patients suggests the need to use molecular-biological diagnostic methods to identify HBV, as well as to analyze the HBV drug resistance mutation before starting antiretroviral therapy for HIV.

Correlation of HBV genotyping and drug resistance mutation in Hohhot, Inner Mongolia

Objective To investigate the relationship between HBV-DNA load and serum markers in chronic hepatitis B(CHB) patients in Hohhot, Inner Mongolia, and to explore the mutation of HBV genotype and nucleoside analogue. Methods From January 2015 to December 2017, one hundred and ninety-three CHB patients hospitalized in the People's Hospital of Inner Mongolia were selected randomly.The clinical diagnostic criteria for all admitted patients were based on the Guidelines for the Prevention and Treatment of Chronic Hepatitis jointly formulated by the Infectious Diseases Society of 2010.The HBV-DNA load of HBV was detected by real-time quantitative PCR, and the correlation between HBV-DNA load and serum markers was analyzed.Seventy-nine patients were selected from 193 hospitalized patients, PCR-reverse dot blot hybridization was used to analyze HBV genotyping and the drug resistance mutations of different genotypes. Results The differences of HBeAb level and HBV-DNA load between HBeAg positive patients and negative patients were statistically significant(all P<0.001). Of 79 serum specimens of HBV infected people, 9 cases(11.4%) were B genotypes, and 70 cases of C genotype(88.6%). Of them, 25 cases had different loci variation, the rate of variation was 31.6%(25/79), with the unit point rtS213T mutation dominated, accounting for about 24.0%(6/25). Conclusion In Hohhot Inner Mongolia patients with CHB, HBV-DNA load with HBeAg and HBe Ab level are correlated; genotype in patients including B type and C type, which is mainly genotype C; patients with CHB mainly had drug resistance to lamivudine and adefovir dipivoxil, mutations including rtS213T, and hybrid mutation. Key words: Hepatitis B, chronic; Hepatitis B virus; Genes, viral; Deoxyribonucleic acid; Hepatitis B e antigens; Hepatitis B surface antigens; Hepatitis B core antigens; Genotype; Drug resistance mutation

Patterns of hepatitis B virus S gene escape mutants and reverse transcriptase mutations among genotype D isolates in Jordan.

BackgroundHepatitis B virus (HBV) is an important infectious cause of morbidity and mortality in Jordan. HBV genotype D is the most prevalent in the country. Virus escape mutants in the HBV S region is an important public health problem halting preventive efforts. The aim of the current study was to investigate patterns of HBV escape and resistance mutations and to assess domestic transmission of the virus.MethodsPatients infected with HBV were recruited at Jordan University Hospital (n = 56) and were diagnosed during (1984–2012). A total of 37 partial HBV S sequences were generated using Sanger’s method. Mutation analysis was done using the HIV grade HBV drug resistance interpretation online tool and Geno2pheno (HBV) online tools. Domestic transmission of HBV was assessed using maximum likelihood phylogenetic inference with similar GenBank sequences.ResultsGenotyping revealed an exclusive presence of sub-genotype D1. Typical HBV escape mutants were identified in seven patients. These mutations included: L109R, Q129R, M133L, S143L and D144E with overall prevalence of 18.9% (95% CI [9.5–34.2]). Reverse transcriptase (RT) sequence analysis showed mutations in three patients with overall prevalence of 8.1% (95% CI [2.8–21.3]). RT mutations included: V173L, S202I, L180M, M204V and T184A. Transmission cluster analysis revealed a relatively high proportion of infections taking place as a result of domestic spread (29.7%).ConclusionsBased on our findings, RT mutation analysis appears to be of high value before the initiation of therapy in patients with chronic HBV infection in Jordan. Phylogenetic analyses revealed a considerable proportion of local spread in the country which should be considered in the preventive infection control efforts.

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.

Correlation of oxidative stress in patients with HBV-induced liver disease with HBV genotypes and drug resistance mutations

ObjectiveThis study aims to explore the correlation of oxidative stress (OxS) in patients with chronic hepatitis B (CHB) and the disease severity with HBV genotypes and drug resistance mutations. MethodsA total of 296 patients with CHB were enrolled into the study. PCR-reverse dot-blot hybridization was used to detect the HBV genotypes (B, C, and D) and the drug resistance-causing HBV mutant genes. In addition, the total oxidative stress (TOS) and total antioxidant status (TAS) were determined, and oxidative stress index (OSI) was calculated and compared. ResultsSerum levels of TOS and OSI, the B/C ratio, and drug resistance mutation rate were increased along with the elevated disease severity degree (CHB<HBC<HCC) (P < 0.05), while the serum TAS level showed the opposite trend (though there was no statistical difference between patients at the CHB and HBC stages). Moreover, patients with drug resistance-causing HBV mutation had higher serum TOS and OSI levels, while lower serum TAS levels (P < 0.05), compared with patients without mutations. Furthermore, mutation site numbers were positively correlated with the disease severity degree (γ = 0.614, P < 0.001). ConclusionThere is oxidative damage in patients with HBV-induced liver disease, and the damage degree is correlated with the HBV genotype and drug resistance mutation. Oxidative stress might be a useful indicator of the progression of HBV-induced liver disease in patients.

Monitoring by high-sensitivity HBV DNA assay during treatment in chronic hepatitis B e antigen negative patients

Objective: To explore the efficacy of tenofovir disoproxil and adefovir dipivoxil treatment in patients with hepatitis B virus e antigen (HBeAg) negative was analyzed through the comparison of highly sensitive HBV viral load monitoring with HBV genotyping and drug resistance mutations. Methods: The clinical data of newly diagnosed chronic hepatitis B patients from January 2015 to June 2017 in outpatients and inpatients were randomly divided into tenofovir and adefovir group. Quantitative detection of HBV DNA levels before therapy and at 12, 24, 48, 96, and 120 weeks after therapy were determined for HBV genotypes and drug-resistant mutations in HBeAg-negative patients. Student's t-test was used to compare the measurement data between groups. The data of comparison between groups were tested by χ (2). Results: A total of 106 cases of HBeAg-negative patients were collected. Tenofovir disoproxil had a higher rate of HBV DNA suppression (54%) than adefovir dipivoxil treatment (42%), but the difference was not statistically significant (P = 0.19). After 120 weeks of treatment, a total of 46 patients (93.9%) were enrolled in the tenofovir disoproxil group with HBV DNA quantitation < 2 000 IU / ml. Adefovir dipivoxil group of patients with HBV DNA < 2 000 IU / ml a total of 40 cases, accounting for 75.5%. The difference between the two groups was statistically significant (P < 0.05). For 49 cases of HBeAg-negative patients, HBV B, C, B and C were mixed before tenofovir dipivoxil treatment, and C1653T, A1762T and G1764A mutation sites were detected in patients with D genotype. Patients C, B, C, B, and C were examined for C1673T, G1896, G1858, G1899A. After treatment, the detection rate of the above mutation sites decreased, but C1653T, C1673T and G1899A were not detected. New mutation sites such as G1915A / C, L180M, M204V, V207I / L, T184A and V173L were detected, Low resistance rate (25%). Conclusion: Tenofovir disoproxil can be recommended as a treatment for HBeAg-negative patients. For HBeAg-negative patients, the choice of high-sensitivity detection of HBV DNA levels, better monitoring of anti-HBV efficacy.

Clinical effect of 156-week telbivudine sequential therapy in HBeAg-positive chronic hepatitis B patients with suboptimal response to pegylated interferon-α-2a therapy

Objective: To investigate the clinical effect of 156-week telbivudine sequential therapy in HBeAg-positive chronic hepatitis B patients with suboptimal response to pegylated interferon-α-2a (Peg-IFN-α-2a) therapy. Methods: A total of 35 HBeAg-positive CHB patients with HBV DNA < 500 IU/ml who were treated with Peg-IFN-α-2a for 48 weeks and did not experience seroconversion of HBeAg were given telbivudine sequential therapy for 156 weeks. HBeAg clearance rate, HBeAg seroconversion rate, HBV DNA clearance rate, safety, and drug resistance rate were analyzed. The t-test was used for the analysis of continuous data and the chi-square test was used for the analysis of categorical data. A multivariate Cox regression analysis was used to identify the influencing factors for HBeAg seroconversion. Results: Telbivudine sequential therapy achieved an ideal HBeAg seroconversion rate of 87.88% with good tolerability and low drug resistance. The HBeAg clearance rate and HBeAg seroconversion rate increased over the time of treatment and were 45.45% and 45.45%, respectively, at 24 weeks and 93.94% and 87.88%, respectively, at 156 weeks. No patient had virologic breakthrough or HBsAg clearance during treatment. The multivariate Cox regression analysis showed that baseline HBsAg level (hazard ratio [HR] = 0.404, P = 0.003) and > 0.5 lg IU/ml reduction in HBeAg at 24 weeks (HR = 2.196, P = 0.048) were predictive factors for HBeAg seroconversion at 156 weeks. Conclusions: In HBeAg-positive CHB patients with suboptimal response to Peg-IFN-α-2a therapy, 156-week telbivudine sequential therapy has a good clinical effect and can be used as an optimal regimen for such patients.

Mutations associated with drug resistance and prevalence of vaccine escape mutations in patients with chronic hepatitis B infection.

The Brazilian public health system (SUS) has provided antiviral drugs for chronic hepatitis B treatment for over 10 years, but a system for monitoring for drug-related resistance mutations is not available. Determine the presence of HBV mutations associated with resistance to nucleos(t)ide analogs among 81 patients with chronic HBV infection in Salvador-BA-Brazil. HBV-DNA was PCR amplified with primers deduced from the rt domain at the HBV P gene, the sequence extended 1032 bp (from amino acid 1 to 344-rt domain). Those sequences were submitted to the HBV drug resistance database to retrieve each mutation according to the genotype. HBV genotype A1 (85.2%) was the most prevalent, followed by genotype A2 (4.9%), F (6.2%), and C1, D2, and D4 (1.2% each). Six patients (7%) exhibited resistance mutations to LAM, ETV, and TDF: two with patterns L180M + M204V and four with other different patterns: L80I + L180M + M204I; L80V + L180M + M204V; M204I; A194T. All of these mutations were present in patients with genotype A (four A1 and two A2). In addition, four mutations in gene S (three cases with the sI195M mutation and one with the W196L mutation), were detected, corresponding to a rate of 6% of vaccine escape mutations. Althougth the small sample size, an association was found between the occurrence of HBV resistance mutations and HBeAg positivity, co-infection with HIV and a history of treatment for HBV and/or HIV.

HIV Therapy without HBV Co-management in Ethiopia Fosters Emergence of Unintended HBV Drug Resistance and Vaccine Evasive Variants

HIV Therapy without HBV Co-management in Ethiopia Fosters Emergence of Unintended HBV Drug Resistance and Vaccine Evasive Variants

Correction: HBV Genotypes and Drug Resistance Mutations in Antiretroviral Treatment-Naive and Treatment-Experienced HBV–HIV-Coinfected Patients

Correction: HBV Genotypes and Drug Resistance Mutations in Antiretroviral Treatment-Naive and Treatment-Experienced HBV–HIV-Coinfected Patients

HBV genotypes and drug resistance mutations in antiretroviral treatment-naive and treatment-experienced HBV-HIV-coinfected patients.

The presence of HBV resistance mutations upon initiation or during antiretroviral therapy (ART) in HIV-coinfected patients is an important determinant of treatment response. The main objective of the study was to determine the prevalence of HBV resistance mutations in antiretroviral treatment-naive and treatment-experienced HBV-HIV-coinfected Ghanaian patients with detectable HBV viraemia. HBV-HIV-coinfected patients who were ART-naive or had received at least 9 months of lamivudine (3TC)-containing ART were enrolled in a cross-sectional study. Demographic and clinical data were collected and HBV DNA quantified. Partial HBV sequences were amplified by PCR and sequenced bi-directionally to obtain a 2.1-2.2 kb fragment for phylogenetic analysis of HBV genotypes and evaluation of drug resistance mutations. Of the 100 HBV-HIV-coinfected study patients, 75 were successfully PCR-amplified, and 63 were successfully sequenced. Of these 63 patients, 27 (42.9%) were ART-experienced and 58 (92.1%) had HBV genotype E. No resistance mutations were observed in the 36 ART-naive patients, while 21 (77.8%) of 27 treatment-experienced patients had resistance mutations. All patients with resistance mutations had no tenofovir in their regimens, and 80% of them had HIV RNA <40 copies/ml. The 3TC resistance mutations rtL180M and rtM204V were observed in 10 (47.6%) of the 21 patients, while 5 patients (23.8%) had rtV173L, rtL180M and rtM204V mutations. A high proportion of HBV-HIV-coinfected patients with detectable viraemia on 3TC-containing ART had resistance mutations despite good ART adherence as determined by HIV RNA suppression. This study emphasizes the need for dual therapy as part of a fully suppressive ART in all HBV-HIV-coinfected patients in Ghana.

Efficacy of 104-week sequential therapy with telbivudine or entecavir in HBeAg-positive chronic hepatitis B patients with suboptimal responses to 24-week therapy with pegylated interferon-α-2a

To investigate the efficacy and safety of 104-week sequential therapy with telbivudine or entecavir in HBeAg-positive chronic hepatitis B (CHB) patients with suboptimal responses to 24-week pegylated interferon-α-2a (PEG-IFN-α-2a) therapy. A total of 130 HBeAg-positive CHB patients with HBV DNA≥5.0 lg IU/ml and a reduction in HBsAg quantitation < 1 lg IU/ml compared with baseline who received PEG-IFN-α-2a therapy for 24 weeks were enrolled and randomly divided into telbivudine group and entecavir group, and 5 of them were lost. HBeAg clearance rate and seroconversion rate, HBV DNA clearance rate, safety, and drug resistance rate at week 104 were observed. The t-test, chi-square test, or multivariate Cox regression analysis were used for statistical analysis of different types of data. At week 104 of treatment, HBV DNA clearance rate showed no significant difference between the telbivudine group and entecavir group (P = 0.363), and the telbivudine group had significantly higher HBeAg clearance rate and HBeAg seroconversion rate than the entecavir group (HBeAg clearance rate: 61.29% vs 23.81%, P < 0.01; HBeAg seroconversion rate: 51.61% vs 19.05%, P < 0.01). Male sex and telbivudine therapy were baseline predictors of HBeAg seroconversion. The multivariate Cox regression analysis (Forward LR, a = 0.05) showed that the presence or absence of HBeAg seroconversion at week 104 was significantly associated with male sex (HR = 4.917), a reduction in HBsAg > 0.5 lg IU/ml at week 12 of treatment compared baseline (HR = 3.514), and a reduction in HBeAg > 1 lg COI at week 12 of treatment compared baseline (HR = 8.651). In HBeAg-positive CHB patients with suboptimal responses to 24-week PEG-IFNα-2a therapy, the sequential therapy with telbivudine helps achieve better HBeAg clearance rate and seroconversion rate compared with the sequential therapy with entecavir and can be used as a therapeutic regimen for such patients. A reduction in HBeAg > 1 lg COI at week 12 of treatment compared baseline can be used as a predictive factor for HBeAg seroconversion at week 104.

HIV therapy with unknown HBV status is responsible for higher rate of HBV genome variability in Ethiopia.

In Ethiopia, HBV and HIV are co-circulating. Since patients are not routinely tested for HBV, the use of antiretroviral drugs could contribute to unintended HBV drug resistance and surface gene variability during HIV coinfection. A total of 161 hepatitis B surface antigen (HBsAg)-positive sera from 58 HIV-coinfected and 103 drug-naive HBV-monoinfected individuals were characterized for HBV drug resistance and immune escape HBsAg variants. HBV polymerase/surface gene fragment of 716 bp was analysed by direct sequencing. In 34 out of 161 study subjects (21.1%) HBV drug resistance mutations (DRMs) were detected with a frequency of 3.1% rtL80F/I, 0.6% rtA181V, 1.2% rtT184S, 6.2% rtV173L, 10.6% rtL180M, 10.6% rtM204V/I and 8.1% rtI233V. The prevalence of the major DRMs in HBV-HIV-coinfected individuals was significantly higher than monoinfected individuals (41.4% versus 10.7%). Lamivudine selected DRMs, that is, rtL180M (29.3%) and rtM204V/I (29.3%) and rtV173L (15.5%) were more prevalent in HBV-HIV-coinfected individuals but absent in HBV-monoinfected individuals. Despite the finding that rtL180M and rtM204V/I were higher among ART-experienced individuals, the overall prevalence of DRMs (48.0% versus 36.4%) showed no significance difference among antiretroviral therapy (ART) status. The study also revealed higher frequency and heterogeneity of putative and known immune escape HBsAg mutations both in the major hydrophilic region (MHR; 68.3%) and outside the MHR (82.5%) of the surface gene. In particular, the 'a' determinant surface gene mutations (sT125S, sA128V, sQ129H/R, sT131I, sC137S, sT143M, sD144D/E, sG145R, sT148P) and the majority of clustered/multiple as well as drug selected immune escape HBsAg mutations were more prevalent in HBV-HIV-coinfected individuals. HIV therapy without HBV co-management in Ethiopia fosters emergence and circulation of HBV variants of public health importance. It is highly recommended to include HBV testing and co-management as part of routine HIV care programmes for a better ART selection.

Response to potent anti-HBV agents in chronic hepatitis B and combined effect of HBV reverse transcriptase mutations

Background and aimResponse to nucleos(t)ide analogue therapy against HBV infection depends on a number of factors. One of them is appearance of drug resistance mutations. The present study aimed to investigate the efficacy of ETV and TDF as anti-HBV agents and to analyze the role of HBV-RT mutations in reducing the efficacy of mentioned drugs. Material and methodsSixty nine treatment naïve CHB patients (mean age 33.8±11.9years) were enrolled and treated with ETV or TDF for one year. Complete virological response (CVR) was defined as undetectable serum HBV DNA after 12months of therapy. Amino acid and nucleotide sequence analyses of HBV-RT region were performed using Geno2pheno HBV drug resistance tool. The 3D model of HBV-RT protein was built by I-TASSER server and RMSD was calculated between wild type and mutated HBV-RT protein. ResultsAfter 12months of treatment, four CHB patients did not achieve CVR and all of them were with HBV genotype D. HBeAg seroconversion was achieved in 56% HBeAg positive patients after 12months of antiviral therapy. The HBV-RT amino acid sequences from these four patients were used for in-silico analysis. It was found that the presence of many mutations in HBV-RT region of HBV isolated from these patients led to a high degree of variation in configuration of atoms of HBV-RT protein and also caused displacement of active site of this protein. ConclusionThe efficacy of antiviral drugs in inhibiting HBV replication may be reduced by combined effect of many HBV-RT mutations; however, an in vitro study is needed to validate the findings.