scholarly journals Comparison of Two Diagnostic Methods for the Detection of Hepatitis B Virus Genotypes in the Slovak Republic

Pathogens ◽  
2021 ◽  
Vol 11 (1) ◽  
pp. 20
Author(s):  
Mariia Logoida ◽  
Pavol Kristian ◽  
Andrea Schreiberova ◽  
Patrícia Denisa Lenártová ◽  
Veronika Bednárová ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Sanger Sequencing ◽  
Slovak Republic ◽  
Diagnostic Methods ◽  
Response To Treatment ◽  
Health Concern ◽  
Rt Pcr ◽  
B Virus ◽  
Commercial Kit

The hepatitis B virus (HBV), belonging to the Hepadnaviridae family, is responsible for a global health concern still in the 21st century. The virus is divided into 10 genotypes, which differ in geographical distribution and in their effect on disease progression and transmission, susceptibility to mutations, and response to treatment. There are many methods for diagnostics of HBV and differentiating its genotypes. Various commercial kits based on real-time polymerase chain reaction (RT PCR) and hybridization available, as well as whole genome sequencing or the sequencing of only individual parts of the genomes. We compared a commercial kit AmpliSens HBV-genotype-FRT, based on RT PCR, with an adapted method of amplification of the surface genomic region combined with Sanger sequencing. In the examined samples we identified the A, B, C, D, and E genotypes. By PCR with Sanger sequencing, the genotypes were determined in all 103 samples, while by using the commercial kit we successfully genotyped only 95 samples, including combined genotypes, which we could not detect by sequencing.

scholarly journals Development and evaluation of an in-house ELISA to detect hepatitis B virus surface antigen in resource-limited settings

2014 ◽  
Vol 39 (2) ◽  
pp. 65-68 ◽  
Author(s):  
K Fatema ◽  
S Tabassum ◽  
A Nessa ◽  
M Jahan
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Surface Antigen ◽  
Hbv Infection ◽  
Health Concern ◽  
Enzyme Linked Immunosorbent Assay ◽  
Virus Surface ◽  
B Virus ◽  
Commercial Kits ◽  
Checkerboard Titration

Hepatitis B virus (HBV) infection is of global public health concern. Among various serological tests used for the diagnosis and screening of HBV infection, the enzyme-linked immunosorbent assay (ELISA) to detect hepatitis B surface antigen (HbsAg) is most widely used. The present study was designed to develop and standardize a cost effective in-house ELISA for the detection of HbsAg and compare its performance with two established commercial kits. The concentrations of coating antibody, conjugates and sera were fixed by checkerboard titration. Using known HBsAg positive and negative sera, four different concentrations (1, 0.5, 0.25 and 0.125 ?g/well) of coating anti-HBs were applied. Similarly, serial dilutions of patients’ sera (1 in 2, 1 in 3, 1 in 5 and 1 in 9) and conjugates (1 in 2, 1 in 3, 1 in 5, 1 in 9 and 1 in 17) were evaluated by checkerboard titration. The optimal concentration of coating antibody was determined at 0.25 ?g/well and 1 in 9 dilution for both conjugates and sera. The performance comparison of our in-house ELISA showed excellent correlation with two commercial kits (Pearson 0.957, P=0.001 for monoclonal antibody coated kit and Pearson 0.929, P=0.000 for polyclonal antibody coated kit) when OD values were compared. All commercial kit proven positive samples was positive while all negative samples were negative with the in-house ELISA resulting in 100% sensitivity and specificity. The results of our study demonstrated that our inhouse ELISA for detection of HBsAg was equally as sensitive and specific as two well-known commercial kits. Thus, this system may be a useful tool for diagnostic and screening purposes, as well as outbreak investigations. DOI: http://dx.doi.org/10.3329/bmrcb.v39i2.19644 Bangladesh Med Res Counc Bull 2013; 39: 65-68

scholarly journals How to Clear HBV cccDNA: CRISPR/Cas9 Might Be Promising

2017 ◽  
Author(s):  
Yan Qiu ◽  
Ying Liu ◽  
Wen Ren ◽  
Jing Ren
Keyword(s):  
Chronic Hepatitis ◽  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Chronic Hepatitis B ◽  
Health Concern ◽  
Liver Carcinoma ◽  
Major Health ◽  
Circular Dna ◽  
Hbv Cccdna ◽  
B Virus

BACKGROUND: Chronic hepatitis B infected with Hepatitis B virus remains a major health concern worldwide. Despite standard interferon-α and nucleotide analogues have been shown to reduce the deterioration of liver disease among chronic hepatitis B patients, covalently closed circular DNA was still difficult to eradicate. METHODS: A literature search of Pubmed and Web of science was performed with the following key words: ‘CRISPR’, ‘CRISPR/Cas9’, ‘hepatitis B’, ‘HBV’, ‘chronic hepatitis B’ and ‘HBV cccDNA’. The information about CRISPR/Cas9 for the treatment of HBV cccDNA or hepatitis B was reviewed. RESULTS: CRISPR/Cas9 could treat hepatitis B through suppressing or clearing HBV cccDNA with different gRNAs. CONCLUSION: With the emergence of CRISPR/Cas9 (the RNA-guided clustered regulatory interspaced short palindromic repeats, CRISPR) editing technology, clearance of hepatitis B virus and better prevention of liver carcinoma seemed to be possible.

scholarly journals High Hepatitis B Seroprevalence, Low Knowledge, and Poor Attitude towards Hepatitis B Virus Infection among Market Women in Bolgatanga Metropolis in the Upper East Region of Ghana

2020 ◽  
Vol 2020 ◽  
pp. 1-9
Author(s):  
Gideon K. Helegbe ◽  
Faiza Tanko ◽  
Paul A. Aryee ◽  
Setor Aku Lotsu ◽  
Mathias J. A. Asaarik ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Hepatitis B Surface Antigen ◽  
Hbv Infection ◽  
Health Concern ◽  
East Region ◽  
B Virus ◽  
Market Women ◽  
Upper East Region ◽  
Knowledge And Attitude

The Bolgatanga Municipal Health Directorate has reported liver cirrhosis among the first three diseases causing mortality from 2013 to 2015. This implicates hepatitis B virus (HBV) infection considering its high prevalence among blood donors in the Upper East Region of Ghana. However, for a vulnerable group such as market women, there is not much information with regard to the prevalence, knowledge, and attitude towards HBV infection. Thus, this study sought to bridge this gap by determining the seroprevalence, knowledge, and attitude of market women in the Bolgatanga Municipality of Ghana, towards HBV infection. A cross-sectional descriptive study was conducted (from October 2017 to March 2018) among 404 market women using a pretested questionnaire to ascertain the knowledge and attitudes of market women towards HBV infection, while hepatitis B surface Antigen Rapid Diagnostic Test strips were used to screen for the infection. The study revealed that the seroprevalence of hepatitis B among the market women was 15.6%, and majority of the study subjects (>60%) were unaware of HBV infection. Overall, knowledge on and attitude towards HBV infection were low and poor, respectively, with a significantly high number of the market women not wanting infected individuals to be isolated (p=0.049). A high seroprevalence, together with poor attitude and low knowledge levels, as seen in this study is of great public health concern. The study recommends regular HBV screening for market women for prompt treatment and vaccination as well as continuous health education to increase knowledge level and improve the poor attitudes of market women towards HBV infection.

scholarly journals A Sensitive and Rapid Assay for Investigating Vertical Transmission of Hepatitis B Virus via Male Germ Line Using EGFP Vector as Reporter

2008 ◽  
Vol 2008 ◽  
pp. 1-6 ◽  
Author(s):  
Mohamed Morsi M. Ahmed ◽  
Tian-Hua Huang ◽  
Qing-Dong Xie
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Vertical Transmission ◽  
Germ Line ◽  
Dna Recombination ◽  
Green Fluorescence ◽  
Rt Pcr ◽  
Male Germ Line ◽  
Rapid Assay ◽  
B Virus

Hepatitis B virus (HBV) constitutes a serious menace to man. DNA recombination and sequencing, interspecific in vitro fertilization, single-embryo PCR and RT-PCR were employed to establish a sensitive and rapid assay for exploring the vertical transmission of viruses via male germ line. Plasmid pIRES2-EGFP-HBs which expressed enhanced green fluorescent protein as reporter for the expression of hepatitis B virusSgene was successfully constructed and confirmed by PCR, EcoR I and Sal I digestion, and DNA sequencing. After exposure to the plasmid, human spermatozoa were used to fertilize with zona-free hamster ova. Two-cell embryos were collected and classified into group A with green fluorescence and group B without green fluorescence under fluorescence microscope. The results showed that HBs DNA positive bands were detected in the embryos with green fluorescence (PCR and RT-PCR) and positive control (PCR) indicating expression of pIRES2-EGFP-HBs, and not observed in the embryos without green fluorescence and negative controls (PCR and RT-PCR) indicating no pIRES2-EGFP-HBs in the cells. The advantages and application foreground of this assay for study on vertical transmission of viruses such as HCV, HIV, HPV, and SARS via germ line were discussed.

scholarly journals Retrospective analysis of hepatitis B virus chronic infection in 247 patients: clinical stages, response to treatment and poor prognostic factors

2017 ◽  
Vol 21 (4) ◽  
pp. 441-447 ◽  
Author(s):  
Marlone Cunha-Silva ◽  
Fábio R.T. Marinho ◽  
Paulo F. Oliveira ◽  
Tirzah M. Lopes ◽  
Tiago Sevá-Pereira ◽  
...  
Keyword(s):  
Prognostic Factors ◽  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Retrospective Analysis ◽  
Chronic Infection ◽  
Response To Treatment ◽  
B Virus ◽  
Clinical Stages

scholarly journals Serological and Molecular Detection of Hepatitis B virus among patients referred to Kurdistan Center for Hepatology and Gastroenterology in Sulaimani City/Kurdistan Region of Iraq

2020 ◽  
Vol 4 (2) ◽  
pp. 117
Author(s):  
Raz Sirwan Abdulla ◽  
Salih Ahmed Hama
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Hbv Dna ◽  
Molecular Techniques ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
B Virus ◽  
Polymerase Chain ◽  
Pcr Techniques ◽  
Positive Results

Hepatitis B virus infection is caused by the hepatitis B virus, a major global health problem. This infection can lead to chronic conditions, followed by cirrhosis and hepatocellular carcinoma (HCC). The current study was aimed to detect HBV using serological and molecular techniques. During 2019, 300 blood samples were collected from Kurdistan Center for Hepatology and Gastroenterology in Sulaimani city. Enzyme-linked immunosorbent assay (ELISA) and real-time polymerase chain reaction (RT-PCR) techniques were used for the detection of HBsAg and HBV DNA, respectively. Obtained results were revealed that 92 out of 300 tested patients (30.66%) seropositive for HBsAg. Among 92 seropositive patients, 53 were shown positive results for HBV DNA by RT-PCR. Dental clinic visiting and dialysis were among the important risk factors for HBV transmission. The vast majority of positive results were among males. Smokers showed relatively high rates of positive results. One-third of the referred patients who had liver complaints were positive for HBsAg. More than half of the seropositive patients showed RT-PCR positive results. It was concluded that the molecular method (RT-PCR) is more sensitive and gives a more accurate result than serology (ELISA). Therefore, it can be used as a diagnostic tool for HBV detection.

scholarly journals Real Time PCR HBV-DNA Analysis in HBsAg Positive Patients

2015 ◽  
Vol 10 (2) ◽  
pp. 95-99
Author(s):  
Wasila Rahman ◽  
Muhammad Rabiul Hossain ◽  
Arif Ahmed Khan ◽  
Debashish Saha ◽  
SM Mahbubul Alam ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Antiviral Therapy ◽  
Real Time ◽  
Real Time Pcr ◽  
Armed Forces ◽  
Hbv Dna ◽  
Health Concern ◽  
B Virus ◽  
Pcr Method

Introduction: The hepatitis B virus is a global public health concern and leading cause of chronic liver disease in Bangladesh. For the diagnosis and monitoring of treatment of Hepatitis B virus infection, HBV-DNA detection and quantification is now extensively used worldwide.Objectives: The objective of this study was to detect HBV-DNA by real time PCR method in HBsAg positive patients, to compare the results of HBVDNA detection with HBeAg and Anti-HBe and to monitor the response after antiviral therapy in chronic hepatitis B patients and also to observe the intensity of hepatitis B infection in relation to age and sex.Methods: This was a cross sectional type of study conducted in Armed Forces Institute of Pathology (AFIP), Dhaka Cantonment. In this study, 56 sera of HBsAg positive patients were selected who all were subjected to do HBV-DNA (real time PCR) analysis during the period of 29 July to 30 0ctober, 2013.Results: Out of 56 HBsAg positive patients, HBV-DNA was detected in 34 patients. Among these, 8 (23.5%) patients were HBeAg positive, 16 (47%) patients were anti-HBe positive and 10 (29.5%) were negative for both HBeAg and anti-HBe. Age limit of patients was up to 60 years. HBV-DNA positive patients showed male predominance; 26 (76.5%) patients were male and 8 (23.5%) patients were female. Mean age of the patients was 35±14 years. Among 56 HBsAg positive patients, fifteen were receiving antiviral therapy. Out of them, HBV-DNA was decreased among 4 patients and could not be detected among 11 patients.Conclusion: Real time PCR method of detection of HBV-DNA is very important in patients who are HBeAg negative and this method is also applied to monitor treatment response to antivirals and to detect occult HBV infections immune control phase and also to detect reactivation of HBV cases.Journal of Armed Forces Medical College Bangladesh Vol.10(2) 2014

scholarly journals Baruch Blumberg (1925-2011) and the discovery of the Hepatitis B virus

2021 ◽  
pp. 18-21
Author(s):  
Bob Weintraub
Keyword(s):  
Hepatitis B Virus ◽  
Liver Cancer ◽  
Hepatitis B ◽  
Genetic Polymorphisms ◽  
Nobel Prize ◽  
Primary Liver Cancer ◽  
Diagnostic Methods ◽  
Inherited Susceptibility ◽  
B Virus

Baruch Blumberg’s research was designed to gain a better understanding of the role of human genetic polymorphisms in relation to inherited susceptibility to disease. This led to the discovery of the hepatitis B virus, diagnostic methods for viral detection, and a vaccine. The vaccination program has prevented the death of millions from primary liver cancer. It was the first widely used vaccine against cancer. For his work with the hepatitis B virus, Blumberg was honored in 1976 by the award of the Nobel Prize in Medicine or Physiology.

IDENTIFICATION AND MOLECULAR-GENETIC CHARACTERISTICS OF THE HEPATITIS B VIRUS AMONG HIV-INFECTED PATIENTS IN ARKHANGELSK

2019 ◽  
Vol 64 (3) ◽  
pp. 105-111
Author(s):  
Y. V. Ostankova ◽  
A. V. Semenov ◽  
E. B. Zueva ◽  
A. A. Totolian
Keyword(s):  
Drug Resistance ◽  
Viral Load ◽  
Hepatitis B Virus ◽  
Antiretroviral Therapy ◽  
Hepatitis B ◽  
Virological Failure ◽  
Diagnostic Methods ◽  
Genetic Characteristics ◽  
Occult Hbv ◽  
B Virus

Aim. To estimate the prevalence and characterize the hepatitis B virus among HIV-infected patients with virological failure of antiretroviral therapy in Arkhangelsk. Material and methods. HBV markers determinations (HBsAg, anti-HBs IgG, anti-HBcor 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. Results. HBsAg-negative (occult) HBV was detect in 28 (43.8%) HIV-infected 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. Conclusion. 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.

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