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1.
Article in English | WPRIM | ID: wpr-929022

ABSTRACT

OBJECTIVES@#Hepatitis B virus related acute-on-chronic liver failure (HBV-ACLF) is the most common type of liver failure in China, with a high mortality. Early rapid reduction of HBV-DNA load can improve the survival rate of HBV-ACLF patients. At present, the commonly used drugs are nucleoside (acid) analogues, such as entecavir (ETV), tenofovir, and so on. The newly listed tenofovir alafenamide fumarate (TAF) has attracted great attention of clinicians because of its stronger antiviral effect, higher transaminase normalization rate, better bone and kidney safety, and zero drug resistance. However, there are few clinical research data on the efficacy and safety of TAF in the treatment of Chinese HBV-ACLF patients, and there is a lack of pharmacoeconomic evaluation. This study aims to compare the efficacy, safety, and cost-effectiveness between TAF and ETV in patients with HBV-ACLF.@*METHODS@#The data were collected from 196 HBV-ACLF patients (80 patients in the TAF group and 116 patients in the ETV group) who were hospitalized in Xiangya Hospital, Central South University from May 2020 to March 2021. Biochemistry and virology were detected before and after treatment (at baseline, Week 2, 4, and 12). Clinical features, disease prognosis, and cost-effectiveness were compared between the 2 groups. According to the baseline, HBV-ACLF patients were divided into 4 stages including pre-liver failure stage, early stage, medium stage, and end stage. And the liver transplantation rate and mortality was also compared. Pharmacoeconomic evaluation was taken using cost-effectiveness analysis and cost minimization analysis..@*RESULTS@#After 4 weeks of treatment, there were no significant differences in the efficacy (liver function, viral load) between the 2 groups (all P>0.05). The TAF group showed lower creatinine [(80.35±18.77) μmol/L vs (105.59±82.32) μmol/L, P<0.05] and higher estimated glomerular filtration rate (eGFR) levels [(95.65±23.21) mL/(min·1.73 m2) vs (82.68±26.32) mL/(min·1.73 m2), P<0.05] than the ETV group. After 12 weeks of treatment, the analysis of overall the liver transplantation rate and mortality between the 2 groups showed similar conclusion. However, the TAF group had a lower the liver transplantation rate and mortality than the ETV group in patients with pre-liver failure (0vs13.89%, P<0.05). No evident distinction was found in the liver transplantation rate and mortality during the early, medium, or end stages of liver failure (13.04% vs 17.65%, 37.50% vs 37.04%, and 54.55% vs 68.42%, respectively). Ratio of cost to effectiveness in the ETV group was higher than that in the TAF group.@*CONCLUSIONS@#TAF is not more efficient than ETV group in improving liver function and reducing viral load for HBV-ACLF patients and they also show similar safety. However, TAF has a greater advantage over ETV not only in preserving renal function, but also in reducing the liver transplantation rate and mortality in patients with pre-liver failure. TAF can provide economic benefit to patients with HBV-ACLF.


Subject(s)
Acute-On-Chronic Liver Failure/drug therapy , Alanine/therapeutic use , Antiviral Agents/therapeutic use , Guanine/analogs & derivatives , Hepatitis B virus/genetics , Hepatitis B, Chronic/drug therapy , Humans , Tenofovir/analogs & derivatives , Treatment Outcome
2.
Article in English | WPRIM | ID: wpr-928926

ABSTRACT

OBJECTIVE@#To compare the circular pathological changes of chronic hepatitis B (CHB) patients according to the tongue diagnosis.@*METHODS@#Totally 41 CHB patients with typical white tongue coating (WTC) or yellow tongue coating (YTC) were enrolled and 14 healthy volunteers with normal tongue manifestation served as controls. The mRNA expression of peripheral leukocytes was detected by GeneChips, and 9 genes were randomly selected for expression validation. Circular metabolites were detected by gas chromatographymass spectrometry. Biological information was analyzed based on ingenuity pathways analysis or metabolomics database and the integrated networks were constructed by ClueGO.@*RESULTS@#A total of 945 and 716 differentially expressed genes were found in patients with WTC and YTC relative to healthy volunteers respectively. The biological information analysis indicated that CHB patients had obviously increased functions in cell death, apoptosis and necrosis (Z-score ⩾2, P<0.05) and decreased activation in T lymphocytes (Z-score ⩽-2, P<0.05), regardless of the tongue manifestation. Compared to patients with WTC, the YTC patients were predicted to be more active in functions related to virus replication (Z-score ⩾2, P<0.05), and the content of circular fatty acids, such as oleic acid (P=0.098) and lauric acid (P=0.035), and citric acid cycle-related metabolites were higher in the YTC patients (P<0.1). The integrated analysis based on differential genes and metabolites indicated that the most difference in the biological function network between the WTC and YTC patients was tumor necrosis factor receptor associated factor 6 mediated-nuclear factor kappa-B activation process.@*CONCLUSIONS@#CHB patients with YTC had more severe inflammation and fatty acids metabolism aberrant than patients with WTC. The results facilitate the modern pathological annotation of Chinese medicine tongue diagnosis theory and provide a reference for the interpretation of pharmacological mechanisms of Chinese medicine treatment.


Subject(s)
Fatty Acids , Hepatitis B virus/genetics , Hepatitis B, Chronic , Humans , Metabolomics , T-Lymphocytes , Tongue
3.
Article in Chinese | WPRIM | ID: wpr-928704

ABSTRACT

OBJECTIVE@#To evaluate the risk of reentry in HBV reactive blood donors and feasibility of HBV reentry strategy.@*METHODS@#HBsAg+ or HBV DNA+ donors who had been quarantined for more than 6 months in Jiangsu Province could propose for reentry application. Blood samples were routinely screened by dual-ELISA for HBsAg, anti-HCV, HIV Ab/Ag, and anti- Treponema pallidum and those non-reactive ones were tested by minipool nucleic acid testing (NAT) for three times. To identify occult HBV donors, samples of NAT non-reactive were further tested by electrochemiluminescence immunoassay (ECLIA) for HBV seromarkers (including HBsAg, HBsAb, HBeAg, HBeAb, and HBcAb). Donors of only 4 ECLIA patterns were accepted to reentry, including all 5 HBV seromarkers negative, anti-HBs only but having history of hepatitis B vaccine injection, HBcAb only, HBsAb+ / HBcAb+ with HBsAb more than 200 IU/L. Additionally, the detection rate of HBV infection was compared between routine screening mode and ECLIA, as well as the reentry qualified rate of HBsAg+ and HBV DNA+ blood donors.@*RESULTS@#From Oct. 2016 to Aug. 2019, a total of 737 HBV reactive donors had applied for reentry, including 667 HBsAg+ reactive and 70 HBV DNA+ reactive donors. Among 3 screening methods, the highest HBV detection rate (43.15%, 318/737) was observed on ECLIA, while only 4.75% (35/737) on ELISA and 3.12% (23/737) on NAT, respectively. Among 4 qualified patterns of HBV serological markers, the highest proportion was found in the all negative group (22.90%, 155/677), followed by the group with HBsAb+ only and history of hepatitis B vaccine injection (19.35%, 131/677), and the median concentration of HBsAb was 237.7 IU/L. The unqualified rate of HBV DNA+ donors was 82.86%, which was significantly higher than 47.98% of HBsAg+ donors.@*CONCLUSION@#Routine screening tests merely based on ELISA and NAT could miss occult HBV donors and may not be sufficient for blood safety. HBsAb concentration and vaccine injection history should be included in the evaluation of HBV reactive donors who intend to apply for reentry. There is a relatively larger residual risk of occult HBV infection in blood donors quarantined for HBV DNA reactive.


Subject(s)
Blood Donors , DNA, Viral , Hepatitis B , Hepatitis B Surface Antigens , Hepatitis B virus/genetics , Humans
4.
Chinese Journal of Hepatology ; (12): 429-438, 2022.
Article in Chinese | WPRIM | ID: wpr-928467

ABSTRACT

Hepatitis B virus (HBV) infection remains to be the major cause of chronic liver diseases in China. Since the nucleos(t)ide analogues and pegylated interferon-alpha do not directly target the covalently closed circular DNA (cccDNA) in the nuclei of HBV-infected hepatocytes, those standard-of-care medications cannot efficiently cure the infected hepatocytes and rarely achieve the functional cure of chronic hepatitis B (CHB). Therefore, new antiviral drugs targeting distinct steps of HBV replication and immunotherapeutics reinvigorating antiviral immune responses are urgently needed for the functional cure of CHB. Based on the extensive discussion of the biological and clinical significance of new virologic biomarkers and distinct mechanism of drug candidates currently in clinical development, we propose that the selection of virologic and immunological biomarkers for evaluation of therapeutic efficacy as well as setting the therapeutic endpoints in the clinical trials should be based on the mode of action of investigational drugs. In addition, due to the complexity of CHB pathogenesis, selection of specific subpopulation of CHB patients for the clinical trials of drugs with a specific mode of action should also be considered.


Subject(s)
Antiviral Agents/therapeutic use , Biomarkers , DNA, Circular , DNA, Viral , Hepatitis B/drug therapy , Hepatitis B Surface Antigens , Hepatitis B virus/genetics , Hepatitis B, Chronic , Humans , Virus Replication
5.
Chinese Journal of Biotechnology ; (12): 893-902, 2022.
Article in Chinese | WPRIM | ID: wpr-927752

ABSTRACT

Hepatitis B virus (HBV) infection is one of the most serious public health problems. HBV infection could lead to hepatitis B, and even further develop into hepatic cirrhosis and hepatocellular carcinoma. Interferon lambda (IFN-λ) is a member of the interferon (IFN) family and an important cytokine for antiviral defense. There are four members in IFN-λ family, including IFN-λ1, IFN-λ2, IFN-λ3, and IFN-λ4. The genetic polymorphisms in the IFN-λ genes are associated with HBV replication and treatment response of HBV patients. In this review, we summarized the roles of genetic polymorphisms of the IFN-λ genes played in HBV infection, disease progression and treatment, with the aim to better understand their function. This review could serve as a reference for the HBV prevention and treatment of HBV patients, as well as for future clinical usage.


Subject(s)
Antiviral Agents/pharmacology , Hepatitis B/genetics , Hepatitis B virus/genetics , Humans , Interferons/pharmacology , Liver Neoplasms , Polymorphism, Genetic , Virus Replication/genetics
6.
Braz. j. biol ; 82: e245813, 2022. tab, graf
Article in English | LILACS, VETINDEX | ID: biblio-1285592

ABSTRACT

Abstract Hepatitis B virus infection is perilous among the five types of Hepatitis, as it remains clinically asymptomatic. The present study draws up-to-date prevalence of Hepatitis B virus (HBV) in the general population of Mardan, Khyber Pakhtunkhwa Pakistan. The blood samples from 4803 individuals including 2399 male and 2404 females were investigated. All the suspected samples were analyzed for hepatitis B surface antigen using Immuno-chromatographic test (ICT), Enzyme-linked immunosorbent assay (ELISA), and followed by Reverse transcription-polymerase chain reaction (RT-PCR). Results showed that 312 (13.00%) out of 2399 individuals contained antibodies in their blood against HBV, while among the different age groups, the highest incidences of HBV antibodies were found in the age of 21-30 groups (10.73%). Furthermore, the ICT positive samples were screened by nested polymerase chain reaction to detect the existence of active HBV-DNA. It was observed that 169 (7.04%) out of (2399) male of the total population (4803) tested was positive. On the other hand, the female 463 (19.25%) possessed antibodies in their blood against HBV. Accumulatively, our results showed a higher percentage of HBV prevalence in males than females in the age group 21-30 years. The total HCV infected in Mardan general population was recorded at 5.7% comprising both male and female.


Resumo A infecção pelo vírus da hepatite B é perigosa entre os cinco tipos de hepatite, pois permanece clinicamente assintomática. O presente estudo traça a prevalência atualizada do vírus da hepatite B (HBV) na população geral de Mardan, Khyber Pakhtunkhwa, no Paquistão. Amostras de sangue de 4.803 indivíduos, incluindo 2.399 homens e 2.404 mulheres, foram investigadas. Todas as amostras suspeitas foram analisadas para o antígeno de superfície da hepatite B usando teste imunocromatográfico (ICT), enzyme-linked immunosorbent assay (ELISA), seguido por transcrição reversa-reação em cadeia da polimerase (RT-PCR). Os resultados mostraram que 312 (13,00%) de 2.399 indivíduos continham anticorpos no sangue contra o VHB, enquanto, entre as diferentes faixas etárias, as maiores incidências de anticorpos VHB foram encontradas nos grupos de 21 a 30 anos (10,73%). Além disso, amostras positivas para ICT foram rastreadas por reação em cadeia da polimerase aninhada para detectar a existência de HBV-DNA ativo. Observou-se que 169 (7,04%) de 2.399 homens do total da população (4803) testados foram positivos. Por outro lado, 463 mulheres (19,25%) possuíam anticorpos no sangue contra VHB. Acumulativamente, nossos resultados mostraram uma porcentagem maior de prevalência de HBV em homens do que em mulheres na faixa etária de 21 a 30 anos. O total de HCV infectados na população geral de Mardan foi registrado em 5,7%, incluindo homens e mulheres.


Subject(s)
Humans , Male , Female , Adult , Young Adult , Hepatitis B virus/genetics , Hepatitis B/epidemiology , Pakistan/epidemiology , Prevalence , Hepatitis B Surface Antigens
7.
Gac. méd. Méx ; 157(1): 37-42, ene.-feb. 2021. tab
Article in Spanish | LILACS | ID: biblio-1279071

ABSTRACT

Resumen Introducción: La identificación de portadores del virus de la hepatitis B en donantes de sangre es imperativo para evitar la transmisión de la enfermedad a través de transfusiones sanguíneas. Objetivo: Determinar si los donantes de sangre con resultados positivos de los marcadores serológicos HbsAg y anti-HBc eran portadores de ADN del virus de la hepatitis B. Métodos: Se recolectaron 12 745 muestras de seis bancos de sangre ecuatorianos, las cuales fueron analizadas con pruebas serológicas para identificar los marcadores infecciosos HBsAg, anti-HBc, anti-HBs mediante prueba ELISA automatizada. Todas las muestras positivas para uno, dos o los tres marcadores fueron analizadas con técnica molecular para determinar la presencia de ADN viral. Resultados: Se identificó que 27.5 % de las muestras reactivas solo a anti-HBc y 100 % de las muestras con resultados positivos de HBsAg/anti-HBc-IgM/IgG presentaron ADN del virus de la hepatitis B (p = 0.001). Conclusiones: La elección de los marcadores de infección y los métodos de detección definen los resultados. Es importante la realización de dos pruebas serológicas y una molecular para identificar a los portadores del virus de la hepatitis B y evitar su transmisión.


Abstract Introduction: Identification of hepatitis B virus carriers in blood donors is imperative in order to avoid transmission of the disease via blood transfusion. Objective: To determine if blood donors with positive results for serological markers HBsAg and anti-HBc were hepatitis B virus DNA carriers. Methods: 12,745 samples were collected from six Ecuadorian blood banks and analyzed for HBsAg, anti-HBc and anti-HBs infectious markers by automated ELISA. All samples that tested positive for one, two or all three markers were analyzed with molecular techniques to determine the presence of viral DNA. Results: 27.5 % of the samples that were reactive for anti-HBc alone and 100 % of those with positive results for HbsAg and IgM/IgG anti-HBc were identified to contain hepatitis B virus DNA (p = 0.001). Conclusions: The selection of infection markers, as well as the detection methods define the results. Performing two serological and one molecular test is important in order to identify hepatitis B virus carriers and prevent its transmission.


Subject(s)
Humans , Blood Donors/statistics & numerical data , DNA, Viral/blood , Immunoglobulin G/blood , Immunoglobulin M/blood , Hepatitis B virus/genetics , Hepatitis B Surface Antigens/blood , Blood Banks , Enzyme-Linked Immunosorbent Assay/methods , Biomarkers/blood , Carrier State/diagnosis , Carrier State/virology , Hepatitis B virus/immunology , Ecuador
8.
Chinese Medical Journal ; (24): 1160-1167, 2021.
Article in English | WPRIM | ID: wpr-878100

ABSTRACT

BACKGROUND@#Hepatitis B core-related antigen (HBcrAg) is a promising disease-monitoring marker for chronic hepatitis B (CHB). We investigated correlations between HBcrAg with antiviral efficacy and virological and histological variables.@*METHODS@#One hundred and forty-five CHB patients from the mainland of China between August 2013 and September 2016 who underwent liver biopsy received entecavir therapy and had paired liver biopsy at 78 weeks. We analyzed correlations between HBcrAg and virological and histological variables in hepatitis B e antigen (HBeAg)-positive and HBeAg-negative patients. We also explored the predictors of HBeAg loss after 78 weeks of antiviral therapy. Pearson correlation analysis and logistic forward stepwise regression were the main statistic methods.@*RESULTS@#HBeAg-positive patients (n = 93) had higher baseline HBcrAg (median 7.4 vs. 5.3 log10 U/mL P < 0.001) and greater HBcrAg declines (median 1.6 vs. 0.9 log10 U/mL P = 0.007) than HBeAg-negative patients after 78 weeks of therapy. At baseline, HBcrAg correlated with hepatitis B virus (HBV) DNA in both HBeAg-positive (r = 0.641, P < 0.001) and -negative patients (r = 0.616, P < 0.001), with hepatitis B surface antigen (HBsAg) in HBeAg-positive patients (r = 0.495, P < 0.001), but not with anti-hepatitis B virus core antibody (anti-HBc). Weak correlations existed between HBcrAg, histology activity index (HAI; r = 0.232, P = 0.025), and Ishak fibrosis score (r = -0.292, P = 0.005) in HBeAg-positive patients. At 78 weeks, significant correlations existed only between HBcrAg and anti-HBc in HBeAg-positive (r = -0.263, P = 0.014) and HBeAg-negative patients (r = -0.291, P = 0.045). Decreased HBcrAg significantly correlated with reduced HBV DNA (r = 0.366, P = 0.001; r = 0.626, P < 0.001) and HBsAg (r = 0.526, P = 0.001; r = 0.289, P = 0.044) in HBeAg-positive and -negative patients, respectively, and with reduced HAI in HBeAg-positive patients (r = 0.329, P = 0.001). Patients with HBeAg loss (n = 29) showed a larger reduction in HBcrAg than those without (median 2.3 vs. 1.3 log10 U/mL, P = 0.001). In multivariate analysis, decreased HBcrAg was an independent predictor of HBeAg loss (P = 0.005).@*CONCLUSIONS@#HBcrAg reflects viral replication and protein production. Decreased HBcrAg could predict HBeAg loss after antiviral therapy.@*TRIAL REGISTRATION@#Clinical Trials.gov: NCT01962155; https://www.clinicaltrials.gov/ct2/show/NCT01962155?term=NCT01962155&draw=2&rank=1.


Subject(s)
Antiviral Agents/therapeutic use , Biomarkers , China , DNA, Viral , Hepatitis B Core Antigens/therapeutic use , Hepatitis B Surface Antigens , Hepatitis B e Antigens , Hepatitis B virus/genetics , Hepatitis B, Chronic/drug therapy , Humans , Virus Replication
9.
Chinese Medical Journal ; (24): 2810-2817, 2021.
Article in English | WPRIM | ID: wpr-921217

ABSTRACT

Low-level viremia (LLV) was defined as persistent or intermittent episodes of detectable hepatitis B virus (HBV) DNA (<2000 IU/mL, detection limit of 10 IU/mL) after 48 weeks of antiviral treatment. Effective antiviral therapies for chronic hepatitis B (CHB) patients, such as entecavir (ETV), tenofovir disoproxil fumarate (TDF), and tenofovir alafenamide (TAF), have been shown to inhibit the replication of HBV DNA and prevent liver-related complications. However, even with long-term antiviral therapy, there are still a number of patients with persistent or intermittent LLV. At present, the research on LLV to address whether adversely affect the clinical outcome is limited, and the follow-up treatment for these patients is open to question. At the same time, the mechanism of LLV is not clear. In this review, we summarize the incidence of LLV, the association between LLV and long-term outcomes, possible mechanisms, and management strategies in these patient populations.


Subject(s)
Antiviral Agents/therapeutic use , DNA, Viral , Hepatitis B virus/genetics , Hepatitis B, Chronic/drug therapy , Humans , Nucleosides/therapeutic use , Tenofovir/therapeutic use , Treatment Outcome , Viremia/drug therapy
10.
Article in English | WPRIM | ID: wpr-879952

ABSTRACT

The pathogenesis of hepatitis B virus (HBV)-associated hepatocellular carcinoma (HCC) is complicated with the crosstalk of multiple factors and the multi-step processes. The main mechanisms underlying the HBV-induced HCC include:①integration of HBV DNA into the host hepatocyte genome to alter gene function at the insertion site,resulting in host genome instability and expression of carcinogenic truncated proteins;②HBV gene mutations at S,C,and X coding regions in the genome;③HBV X gene-encoded HBx protein activates proto-oncogenes and inhibits tumor suppressor genes,leading to the HCC occurrence. In this article,the recent research progress on the molecular mechanism of HBV-induced HCC is comprehensively reviewed,so as to provide insights into the prevention,early prediction and postoperative adjuvant therapy of HCC.


Subject(s)
Carcinoma, Hepatocellular , Hepatitis B/complications , Hepatitis B virus/genetics , Hepatocytes , Humans , Liver Neoplasms
11.
Rev. Soc. Bras. Med. Trop ; 54: e08072020, 2021. graf
Article in English | LILACS | ID: biblio-1340822

ABSTRACT

Abstract INTRODUCTION: Hepatitis B virus (HBV) infection is a public health problem; therefore, we aimed to report HBV genotypes in Ceará, Brazil. METHODS: A total of 103 HBsAg-positive samples were subjected to HBV genotyping and subgenotyping. RESULTS: The following genetic compositions of samples were found: F-54% (F2-83.33%), A-40% (A1-65%), D-6%, C2-1%, E-1%, and G-1%. CONCLUSIONS: Some genotypes are only prevalent in certain parts of the world; however, the State of Ceará is a hub for migration and has one of the most important liver transplantation centers in Brazil, which can explain the prevalence of the F genotype.


Subject(s)
Humans , Gastroenterology , Hepatitis B/epidemiology , Brazil/epidemiology , DNA, Viral/genetics , Hepatitis B virus/genetics , Prevalence , Genotype , Hepatitis B Surface Antigens
12.
Mem. Inst. Oswaldo Cruz ; 115: e200006, 2020. tab, graf
Article in English | LILACS, SES-SP | ID: biblio-1135222

ABSTRACT

BACKGROUND Occult hepatitis B virus (HBV) - characterized by the absence of detectable HBsAg in the presence of HBV DNA - represents a potential threat for blood safety. OBJECTIVES This study was conducted with the aim to investigate the serological and molecular characterization of occult HBV infection (OBI) among blood donors in Mozambique. METHODS 1,502 blood donors were tested for HBsAg. All HBsAg-negative individuals were tested for HBV DNA. Antibodies against HBV core, surface and HBe antigen (anti-HBc, anti-HBs, HBeAg) were measured in HBV DNA positive individuals. FINDINGS 1435 serum samples were HBsAg negative and 16 positive for HBV DNA, 14 confirmed to have OBI, corresponding to a frequency of 0.98%. Of the 14 OBI infections identified, 13/14 (92.8%) were positive for anti-HBc, 4/14 (28.5%) for anti-HBs, and no samples were reactive for HBeAg. Of the 14 OBI cases, nine samples (64.2%) were sequenced for the S/P region. Eight samples (88.9%) belonged to genotype A1 and one (11.1%) to genotype E. One escape mutation (T123A) associated with OBI and various amino acid substitutions for genotype A1 and E were observed. MAIN CONCLUSIONS Our results show the importance of using nucleic acid amplification test to detect occult hepatitis B infection in blood donors in Mozambique.


Subject(s)
Humans , Male , Female , Adult , Blood Donors , Hepatitis B virus/isolation & purification , Hepatitis B virus/genetics , Nucleic Acid Amplification Techniques/methods , Hepatitis B/diagnosis , Hepatitis B Surface Antigens/genetics , Phylogeny , DNA, Viral , Polymerase Chain Reaction , Cross-Sectional Studies , Mozambique
13.
Braz. j. microbiol ; 49(4): 848-855, Oct.-Dec. 2018. tab, graf
Article in English | LILACS | ID: biblio-974300

ABSTRACT

ABSTRACT We studied the role of Thermus thermophilus Recombinase A (RecA) in enhancing the PCR signals of DNA viruses such as Hepatitis B virus (HBV). The RecA gene of a thermophilic eubacterial strain, T. thermophilus, was cloned and hyperexpressed in Escherichia coli. The recombinant RecA protein was purified using a single heat treatment step without the use of any chromatography steps, and the purified protein (>95%) was found to be active. The purified RecA could enhance the polymerase chain reaction (PCR) signals of HBV and improve the detection limit of the HBV diagnosis by real time PCR. The yield of recombinant RecA was ∼35 mg/L, the highest yield reported for a recombinant RecA to date. RecA can be successfully employed to enhance detection sensitivity for the diagnosis of DNA viruses such as HBV, and this methodology could be particularly useful for clinical samples with HBV viral loads of less than 10 IU/mL, which is interesting and novel.


Subject(s)
Bacterial Proteins/genetics , Hepatitis B virus/isolation & purification , Polymerase Chain Reaction/methods , Thermus thermophilus/enzymology , Cloning, Molecular , Recombinases/genetics , Bacterial Proteins/isolation & purification , Bacterial Proteins/metabolism , Gene Expression , Hepatitis B virus/genetics , Polymerase Chain Reaction/instrumentation , Thermus thermophilus/genetics , Recombinases/isolation & purification , Recombinases/metabolism , Escherichia coli/genetics , Escherichia coli/metabolism
14.
Braz. j. infect. dis ; 22(4): 294-304, July-Aug. 2018. tab, graf
Article in English | LILACS | ID: biblio-974222

ABSTRACT

ABSTRACT Background Hepatitis 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. Objective To investigate epidemiological characteristics and HBV genotypes and subgenotypes circulating in a specific city with high HBV prevalence. Methods A 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. Results The 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. Conclusions These 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.


Subject(s)
Humans , Male , Female , Adolescent , Adult , Middle Aged , Young Adult , Hepatitis B virus/genetics , Hepatitis B, Chronic/epidemiology , Drug Resistance, Viral , Antiviral Agents/therapeutic use , Brazil/epidemiology , Hepatitis B virus/drug effects , Polymerase Chain Reaction , Prevalence , Cross-Sectional Studies , Risk Factors , Viral Load , Hepatitis B, Chronic/virology , Genotype , Hepatitis B Surface Antigens/blood , Mutation
15.
Mem. Inst. Oswaldo Cruz ; 113(1): 62-65, Jan. 2018. tab, graf
Article in English | LILACS | ID: biblio-1040579

ABSTRACT

In occult hepatitis B infection (OBI), hepatitis B virus DNA (HBV DNA) can be detected in serum samples; however, oral fluid collection for detection of HBV DNA has not yet been explored, despite the availability of collection devices. Serum and oral fluid samples from 45 hepatitis B core antibody (anti-HBc)-positive patients were collected for the amplification of the HBV polymerase gene. HBV DNA was detected in five serum and four oral fluid samples (the detection limit for oral fluid was 1.656 log IU/mL in paired serum). In conclusion, simple methodologies of sample collection and in-house polymerase chain reaction (PCR) allowed detection of HBV DNA, and these could be used to improve the diagnosis of OBI, especially in locations with limited resources.


Subject(s)
Humans , Male , Female , Adult , Aged , Saliva/virology , DNA, Viral/analysis , Hepatitis B/diagnosis , Hepatitis B Antibodies/analysis , Hepatitis B Surface Antigens/analysis , DNA, Viral/blood , Hepatitis B virus/isolation & purification , Hepatitis B virus/genetics , Polymerase Chain Reaction , Viral Load , Middle Aged
16.
Braz. j. infect. dis ; 21(5): 525-529, Sept.-Oct. 2017. tab
Article in English | LILACS | ID: biblio-888904

ABSTRACT

Abstract Infection by hepatitis B virus (HBV) is a worldwide public health problem. Chronic HBV infection with high viral replication may lead to cirrhosis and/or hepatocellular carcinoma. Mutant HBV strains, such as the HBV A1762T/G1764A double mutant, have been associated with poor prognosis and higher risk of the patient for developing cirrhosis and/or hepatocellular carcinoma. This study analyzed the presence of the HBV A1762T/G1764A double mutant in patients with chronic HBV and its association with clinical parameters such as viral load, aminotransferases, and HBV antigens. A total of 49 patients with chronic hepatitis B were included in the study, and the HBV A1762T/G1764A double mutant strain was detected in four samples (8.16%) by polymerase chain reaction followed by restriction fragment length analysis (PCR-RFLP). The viral load was not significantly different between patients with or without the double mutant strain (p = 0.43). On the other hand, carriers of the HBV A1762T/G1764A double mutant had higher levels of ALT (p = 0.0028), while AST levels did not differ between groups (p = 0.051). In this study, 75% of the samples with the HBV A1762T/G1764A double mutation were HBeAg negative and anti-HBe positive, reflecting seroconversion even though they still displayed high viral loads. Our study has shown that the HBV A1762T/G1764A double mutant strain circulates in Brazilian patients, and is associated with elevated levels of ALT and HBeAg seroconversion.


Subject(s)
Humans , Male , Female , Adult , Middle Aged , Aged , Young Adult , DNA, Viral/genetics , Hepatitis B virus/genetics , Hepatitis B, Chronic/virology , Hepatitis B e Antigens/blood , Mutation/genetics , Brazil , Polymerase Chain Reaction , Cross-Sectional Studies , Sequence Analysis, DNA , Genotype
17.
Mem. Inst. Oswaldo Cruz ; 112(9): 626-631, Sept. 2017. tab
Article in English | LILACS | ID: biblio-894874

ABSTRACT

BACKGROUND In Brazil, few studies have investigated the prevalence of infection with the precore (PC) and basal core promoter (BCP) mutants of the hepatitis B virus (HBV). OBJECTIVES This study aimed to analyse the frequency of PC and BCP mutations among patients infected with HBV and to evaluate the association between the variants and advanced hepatic disease. METHODS A total of 161 patients infected with HBV were studied. To identify PC and BCP mutations, a 501-bp fragment of HBV DNA was amplified and sequenced. FINDINGS PC and BCP regions from HBV strains were successfully amplified and sequenced in 129 and 118 cases, respectively. PC and BCP mutations were detected in 61.0% and 80.6% of the cases, respectively. The A1762T/G1764A variant was identified in 36.7% of the patients with grade 1 and 2 liver fibrosis (29/79) and in 81.8% of the patients with grade 3 and 4 liver fibrosis (9/11) (p < 0.01); in 76.9% of the patients with cirrhosis (10/13) and in 38.1% of the patients without cirrhosis (40/105) (p = 0.01); and in 77.8% of the patients with hepatocellular carcinoma (HCC) (7/9) and in 39.4% of the patients without HCC (43/109) (p = 0.03). MAIN CONCLUSIONS A high prevalence of HBV PC and BCP mutants was found. The A1762T/G1764A variant was independently associated with advanced forms of liver fibrosis, hepatic cirrhosis, and HCC.


Subject(s)
Humans , Female , Adult , Middle Aged , Aged , Aged, 80 and over , Viral Core Proteins/genetics , Hepatitis B virus/genetics , Hepatitis B, Chronic/virology , Liver Cirrhosis/virology , Genotype , Mutation
18.
Braz. j. infect. dis ; 21(4): 424-432, July-Aug. 2017. tab, graf
Article in English | LILACS | ID: biblio-888899

ABSTRACT

Abstract Hepatitis B virus (HBV) is distributed worldwide, with geographical variations regarding prevalence of the different genotypes. The aim of this study was to determine the HBV genotypes and subgenotypes circulating in Southeast Brazil and compare the genetic sequences found with HBV sequences previously described in the world. Sequences from 166 chronic HBV carriers were analyzed using the fragment constituted by 1306 base pairs comprising surface and polymerase regions of the HBV genome. The sequences obtained were submitted to phylogenetic analysis. HBV subgenotypes A1, A2, D1-D4, F2a, and F4 were found. HBV genotype D was the most frequent, found in 99 patients (58.4%). Within this group, subgenotype D3 was the most prevalent, in 73 patients (42.9%). HBV genotype A was identified in 58 (36%) patients, subgenotype A1, in 48 (29.8%) subjects. Genotype F was identified in 9 (5.4%). According to the phylogenetic analysis, the sequences found were grouped with sequences from Europe, Asia and Middle East (subgenotypes D1, D2, D3) and sequences from Latin America and Africa (subgenotype A1). HBV D3 grouped in different clusters inside D3 clade, several of them with sequences isolated in Italy. We also identified eight families whose relatives were infected with the same HBV subgenotype, most with high similarity between sequences. In conclusion, the distribution of the HBV sequences obtained interweaved with sequences from other continents, corresponding to regions from where many immigrants came to this region, in accordance to the hypothesis that the HBV detected over there were brought during the colonization times.


Subject(s)
Humans , Male , Female , Adult , Middle Aged , Aged , Young Adult , Hepatitis B virus/genetics , Hepatitis B, Chronic/virology , Emigrants and Immigrants , Phylogeny , Brazil , DNA, Viral/genetics , Molecular Sequence Data , Sequence Analysis, DNA , Emigration and Immigration , Genotype
19.
Braz. j. med. biol. res ; 50(6): e6050, 2017. tab, graf
Article in English | LILACS | ID: biblio-839310

ABSTRACT

We aimed to investigate the potential role and mechanism of microRNA-30c (miR-30c) in the pathological development of chronic hepatitis B (CHB). The serum levels of miR-30c in hepatitis B virus (HBV) carrier Xinjiang Uygur patients with inactive, low-replicative, high-replicative and HBe antigen-positive CHB were investigated. HepG2 cells were co-transfected with pHBV1.3 and miR-30c mimic or inhibitor or scramble RNA. The effects of miR-30c dysregulation on HBV replication and gene expression, cell proliferation and cell cycle were then investigated. miR-30c was down-regulated in Xinjiang Uygur patients with CHB compared to healthy controls and its expression level discriminated HBV carrier patients with inactive, low-replicative, high-replicative and HBe antigen-positive risk for disease progression. Overexpression of miR-30c significantly inhibited HBV replication and the expressions of HBV pgRNA, capsid-associated virus DNA and Hbx in hepatoma cells. Moreover, overexpression of miR-30c significantly inhibited cell proliferation and delayed G1/S phase transition in hepatoma cells. Opposite effects were obtained after suppression of miR-30c. Our results indicate that miR-30c was down-regulated in Xinjiang Uygur patients with CHB, and miR-30c levels could serve as a marker for risk stratification of HBV infection. Down-regulation of miR-30c may result in the progression of CHB via promoting HBV replication and cell proliferation.


Subject(s)
Humans , Male , Female , Adult , Middle Aged , Disease Progression , Hepatitis B virus/genetics , Hepatitis B, Chronic/blood , MicroRNAs/blood , Carcinoma, Hepatocellular/genetics , Carcinoma, Hepatocellular/pathology , Cell Proliferation/genetics , China , Down-Regulation , Gene Expression Regulation, Viral , Hepatitis B, Chronic/ethnology , Liver Neoplasms/genetics , Liver Neoplasms/pathology , Maze Learning
20.
Article in English | LILACS | ID: biblio-903235

ABSTRACT

ABSTRACT OBJECTIVE To estimate the prevalence of hepatitis B virus and C virus infections and their genotypes and analyze the risk factors for the markers of exposure to hepatitis B virus in female sex workers in a region of intense sex trade. METHODS This is a cross-sectional study performed with four hundred and two female sex workers in Goiânia, Brazil. Data have been collected using the Respondent-Driven Sampling. The women have been interviewed and tested for markers of hepatitis B and C viruses. Positive samples have been genotyped. The data have been analyzed using the Respondent-Driven Sampling Analysis Tool, version 5.3, and Stata 11.0. RESULTS The adjusted prevalence for hepatitis B virus and C virus were 17.1% (95%CI 11.6-23.4) and 0.7% (95%CI 0.1-1.5), respectively. Only 28% (95%CI 21.1-36.4) of the participants had serological evidence of vaccination against hepatitis B virus. Being older (> 40 years), being single, having a history of blood transfusion and use of cocaine, and ignoring the symptoms of sexually transmitted infections were associated with positivity for hepatitis B virus (p < 0.05). We have detected the subgenotype A1 of hepatitis B virus (n = 3) and the subtypes of hepatitis C virus 1a (n = 3) and 1b (n = 1). CONCLUSIONS We can observe a low prevalence of infection of hepatitis B and C viruses in the studied population. However, the findings of the analysis of the risk factors show the need for more investment in prevention programs for sexual and drug-related behavior, as well as more efforts to vaccinate this population against hepatitis B. The genotypes of the hepatitis B virus and C virus identified are consistent with those circulating in Brazil.


Subject(s)
Humans , Male , Female , Adult , Young Adult , Hepatitis B virus/genetics , Hepatitis C/epidemiology , Hepacivirus/genetics , Sex Workers/statistics & numerical data , Hepatitis B/epidemiology , Socioeconomic Factors , Brazil/epidemiology , Sexually Transmitted Diseases , Seroepidemiologic Studies , Prevalence , Cross-Sectional Studies , Risk Factors , Hepatitis C/blood , Genotype , Hepatitis B/blood
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