RESUMEN
The nomenclature of the hepatitis B virus (HBV) genes and their products has developed stepwise, occasionally in an erratic way, creating many misunderstandings, especially among those who do not know the structure of HBV and its genome in detail. One of the most frequent misunderstandings, even presented in leading journals, is the designation of HBV "e"-antigen as envelope or early antigen. Another problem area are the so-called "pre" regions in the HBV genome present upstream of both the core and the surface genes of HBV, inadvertently suggesting that they may be a part of corresponding precursor proteins. Misnomers and misclassifications are frequent in defining the subgenotypes and serological subtypes of HBV. Even the well-established terminology for HBV surface (HBs) or HBV core (HBc) antigen deviates from the conventional virological nomenclature for viral envelopes or capsid proteins/antigens, respectively. Another matter of undesirable variability between publications is the numbering of the nucleotides and the graphical representation of genomic maps. This editorial briefly explains how the nomenclature evolved, what it really means, and suggests how it could be adapted to today's knowledge.
Asunto(s)
Anticuerpos contra la Hepatitis B/inmunología , Antígenos e de la Hepatitis B/inmunología , Virus de la Hepatitis B/inmunología , Hepatitis B/inmunología , Epítopos/genética , Epítopos/inmunología , Variación Genética/genética , Variación Genética/inmunología , Hepatitis B/genética , Hepatitis B/virología , Anticuerpos contra la Hepatitis B/clasificación , Antígenos del Núcleo de la Hepatitis B/clasificación , Antígenos del Núcleo de la Hepatitis B/inmunología , Antígenos de Superficie de la Hepatitis B/clasificación , Antígenos de Superficie de la Hepatitis B/inmunología , Antígenos e de la Hepatitis B/clasificación , Virus de la Hepatitis B/patogenicidad , Humanos , Terminología como AsuntoRESUMEN
BACKGROUND AND OBJECTIVES: Screening of Thai blood donors has resulted in the detection of donors with an occult HBV infection (OBI), where HBsAg is undetectable, but hepatitis B virus (HBV) DNA is present in serum in low concentrations. This study was designed to determine whether the occurrence of OBI in donors was linked to the HBV genotype and possibly to mutations in the surface (S) and core (C) gene regions. MATERIALS AND METHODS: Mutations in the S and C gene regions in 48 Thai donors with OBI were mapped by sequencing. Genotyping was determined with the INNO-LiPA test and by phylogenetic analysis of sequences from the S and C genes. RESULTS: The majority of OBI samples were genotype C (81·3%) with 6·3% of samples being genotype B. In addition, two genotype I isolates were identified. Mutations in the S region (100%) were found especially in loop 1 of the major hydrophilic loop (MHL) at positions I110L, T114S, T126I and S113T, whereas mutations in the C region (65%) were within the basal core promoter region (position A1762T/G1764A) and precore region (position G1896A). CONCLUSION: The majority of OBI samples were HBV genotype C, although genotype I, which is newly emerging in Thailand, was also detected. The study demonstrated that OBI was probably not associated with a particular HBV genotype or with certain mutations in the S and C gene regions. However, mutations in the C gene region which could potentially impair viral replication and HBsAg production and potentially lead to OBI were identified.
Asunto(s)
Oftalmopatías/virología , Virus de la Hepatitis B/genética , Hepatitis B/patología , Secuencia de Bases , Donantes de Sangre , ADN Viral/sangre , Genotipo , Hepatitis B/virología , Antígenos del Núcleo de la Hepatitis B/sangre , Antígenos del Núcleo de la Hepatitis B/clasificación , Antígenos del Núcleo de la Hepatitis B/genética , Antígenos de Superficie de la Hepatitis B/sangre , Antígenos de Superficie de la Hepatitis B/clasificación , Antígenos de Superficie de la Hepatitis B/genética , Humanos , Datos de Secuencia Molecular , Mutación , Filogenia , Reacción en Cadena de la Polimerasa , Regiones Promotoras GenéticasRESUMEN
Vertical transmission from mother to child, the main route of chronic hepatitis B virus (HBV) infection in the East Asia, is considered one of the most important predictors for the response to antiviral therapies as well as its complications such as cirrhosis and hepatocellular carcinoma. Therefore, it is critical in both etiologic and prognostic aspects to confirm whether or not chronic HBV infection is acquired vertically. This study investigated whether mother-to-child infection could be proved by the phylogenetic analyses of HBV pre-S/S genes ever since several decades have elapsed in mother-child pairs with presumed vertical transmission. The pre-S and S regions of HBVs were compared and analyzed phylogenetically in a total of 36 adults (18 mother-child pairs) with chronic HBV infection. All of the isolates of HBV were genotype C and serotype adr. The divergence between mothers and offsprings was 0 to 1.5%. Phylogenetic trees revealed that 17 of 18 pairs (94%) with presumed vertical transmission were grouped into the same cluster. Vertical transmission from mother to child could be strongly suggested even in adults with a history of several decades of HBV infection using the phylogenetic analyses of pre-S and S genes.
Asunto(s)
Antígenos de Superficie de la Hepatitis B/genética , Virus de la Hepatitis B/genética , Hepatitis B Crónica/virología , Adulto , Anciano , ADN Viral/análisis , Femenino , Genotipo , Antígenos de Superficie de la Hepatitis B/clasificación , Virus de la Hepatitis B/clasificación , Virus de la Hepatitis B/metabolismo , Hepatitis B Crónica/diagnóstico , Humanos , Transmisión Vertical de Enfermedad Infecciosa , Masculino , Persona de Mediana Edad , Madres , Filogenia , Reacción en Cadena de la Polimerasa , Análisis de Secuencia de ADN , Serotipificación , Adulto JovenRESUMEN
Vertical transmission from mother to child, the main route of chronic hepatitis B virus (HBV) infection in the East Asia, is considered one of the most important predictors for the response to antiviral therapies as well as its complications such as cirrhosis and hepatocellular carcinoma. Therefore, it is critical in both etiologic and prognostic aspects to confirm whether or not chronic HBV infection is acquired vertically. This study investigated whether mother-to-child infection could be proved by the phylogenetic analyses of HBV pre-S/S genes ever since several decades have elapsed in mother-child pairs with presumed vertical transmission. The pre-S and S regions of HBVs were compared and analyzed phylogenetically in a total of 36 adults (18 mother-child pairs) with chronic HBV infection. All of the isolates of HBV were genotype C and serotype adr. The divergence between mothers and offsprings was 0 to 1.5%. Phylogenetic trees revealed that 17 of 18 pairs (94%) with presumed vertical transmission were grouped into the same cluster. Vertical transmission from mother to child could be strongly suggested even in adults with a history of several decades of HBV infection using the phylogenetic analyses of pre-S and S genes.
Asunto(s)
Adulto , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Adulto Joven , ADN Viral/análisis , Genotipo , Antígenos de Superficie de la Hepatitis B/clasificación , Virus de la Hepatitis B/clasificación , Hepatitis B Crónica/diagnóstico , Transmisión Vertical de Enfermedad Infecciosa , Madres , Filogenia , Reacción en Cadena de la Polimerasa , Análisis de Secuencia de ADN , SerotipificaciónRESUMEN
The detection of HBsAg in blood serum using immune-enzyme analysis techniques decisively matters both for diagnostics of acute and chronic hepatitis B and screening of donor's blood and its components, controls of persons from risk groups of hepatitis B injection. The making of panels containing wide specter of samples of blood serums with sero-variants and mutant forms of surface antigen of hepatitis B virus widespread on the territory of the Russian Federation is necessary to control analytic and diagnostic sensitivity of test systems for detecting HBsAg. The testing of reagents kits to detect HBsAg using twi panels containing recombinant and native variants of HBsAg, demonstrated that these kits enable to detect various sero-vatriants of HBsAg (ayw2, adw2, ayw3varA, ayw3varB, adrq-) in concentration 0.1-0.01 IU/l and the so called elusive mutant forms of HBsAg of recombinant and native origin (G145R, Q129R, Q129H, Q129L, T143K, T126N, T126S, D144A, M133L, K141E and P142S). At that, the sensitivity can differ during the detection of native and recombinant mutant forms. The results testify the importance of using the panels both with recombinant and native samples containing the mutant forms of HBsAg in evaluation of sensitivity of reagents kits.
Asunto(s)
Antígenos de Superficie de la Hepatitis B , Virus de la Hepatitis B , Hepatitis B Crónica , Donantes de Sangre , Antígenos de Superficie de la Hepatitis B/sangre , Antígenos de Superficie de la Hepatitis B/clasificación , Antígenos de Superficie de la Hepatitis B/genética , Virus de la Hepatitis B/clasificación , Virus de la Hepatitis B/inmunología , Hepatitis B Crónica/sangre , Hepatitis B Crónica/diagnóstico , Hepatitis B Crónica/inmunología , Humanos , Mutación , Juego de Reactivos para Diagnóstico , Federación de Rusia , Serotipificación/clasificaciónRESUMEN
AIM: Study heterogeneity ofhepatitis B virus in adult patients with chronic hepatitis B and determination of diagnostic potential of modern test systems with the detection of HBsAg with amino acid substitutions in the main hydrophilic region (MHR). MATERIALS AND METHODS: In 27 hepatitis B virus samples isolated from patients with chronic hepatitis B virus infection living in Vladimir, nucleotide sequence ofgenome region corresponding to preS1/preS2/S genes was determined. RESULTS: In all of the 27 isolates genotype D virus presented by 3 subgenotypes D1, D2, D3 was detected in 18%, 26% and 56% respectively. Based on the distribution of nucleotide substitutions in the compared functional regions of hepatitis B virus (virus entry into the cell coding site (2875 - 2991 n.b.), pre-S2/S promoter region (2994 - 3171 n.b.), 5'-end pre-S2 and S-genes sequences (3172 - 154 n.b. and 155-455 n.b.), MHR (455 - 635 n.b.) and 3'-end S-gene sequence (636 - 835 n.b.), substitutions are mostly concentrated in the promoter region of the S2/S-genes (30.8%). HBsAg serotypes were determined in 24 of 27 cases by using the predicted amino acid sequence, and in 17 cases HBsAg belonged to ayw2 (71%) serotype and in 7 cases - to ayw3 serotype (29%). Amino acid substitutions G145A, M133I, S132T localized in the main hydrophilic region and P217L, S207N, V184A localized in the C-end of the protein C that are connected with diagnostic and vaccine escape were identified in 5 isolates. CONCLUSION: Diagnostic potential of test systems with the detection of HBsAg with known amino acid sequence of the MHR region were studied. Approximately equal potential of 6 test systems to detect HBsAg with amino acid substitutions G145A, M133I and S132T localized in the MHR region were shown.
Asunto(s)
ADN Viral/análisis , Antígenos de Superficie de la Hepatitis B/genética , Virus de la Hepatitis B/genética , Hepatitis B Crónica/diagnóstico , Inmunoensayo , Mutación , Precursores de Proteínas/genética , Adulto , Sustitución de Aminoácidos/inmunología , ADN Viral/biosíntesis , Femenino , Heterogeneidad Genética , Genotipo , Anticuerpos contra la Hepatitis B/análisis , Anticuerpos contra la Hepatitis B/inmunología , Antígenos de Superficie de la Hepatitis B/clasificación , Antígenos de Superficie de la Hepatitis B/inmunología , Virus de la Hepatitis B/clasificación , Virus de la Hepatitis B/inmunología , Hepatitis B Crónica/inmunología , Humanos , Masculino , Persona de Mediana Edad , Sistemas de Lectura Abierta/inmunología , Regiones Promotoras Genéticas/inmunología , Precursores de Proteínas/clasificación , Precursores de Proteínas/inmunología , Estructura Terciaria de Proteína , Federación de Rusia , Sensibilidad y EspecificidadRESUMEN
BACKGROUND AND OBJECTIVES: This study was conducted by the International Consortium for Blood Safety (ICBS) to identify high-quality test kits for detection of hepatitis B virus (HBV) surface antigen (HBsAg) for the benefit of developing countries. MATERIALS AND METHODS: The 70 HBsAg test kits from around the world were evaluated comparatively for their clinical sensitivity, analytical sensitivity, sensitivity to HBV genotypes and HBsAg subtypes, and specificity using 394 (146 clinical, 48 analytical and 200 negative) ICBS Master Panel members of diverse geographical origin comprising the major HBV genotypes A-F and the HBsAg subtypes adw2,4, adr and ayw1-4. RESULTS: Seventeen HBsAg enzyme immunoassay (EIA) kits had high analytical sensitivity <0.13 IU/ml, showed 100% diagnostic sensitivity, and were even sensitive for the various HBV variants tested. An additional six test kits had high sensitivity (<0.13 IU/ml) but missed HBsAg mutants and/or showed reduced sensitivity to certain HBV genotypes. Twenty HBsAg EIA kits were in the sensitivity range of 0.13-1 IU/ml. The other eight EIAs and the 19 rapid assays had analytical sensitivities of 1 to >4 IU/ml. These assays were falsely negative for 1-4 clinical samples and 17 of these test kits showed genotype dependent sensitivity reduction. Analytical sensitivities for HBsAg of >1 IU/ml significantly reduce the length of the HBsAg positive period which renders them less reliable for detecting HBsAg in asymptomatic HBV infections. Reduced sensitivity for HBsAg with genetic diversity of HBV occurred with genotypes/subtypes D/ayw3, E/ayw4, F/adw4 and by S gene mutants. Specificity of the HBsAg assays was >or=99.5% in 57 test kits and 96.4-99.0% in the remaining test kits. CONCLUSION: Diagnostic efficacy of the evaluated HBsAg test kits differed substantially. Laboratories should therefore be aware of the analytical sensitivity for HBsAg and check for the relevant HBV variants circulating in the relevant population.
Asunto(s)
Antígenos de Superficie de la Hepatitis B/sangre , Hepatitis B/epidemiología , Técnicas para Inmunoenzimas/métodos , África/epidemiología , Secuencia de Aminoácidos , Asia/epidemiología , Países en Desarrollo , Reacciones Falso Negativas , Genes Virales , Genotipo , Salud Global , Hepatitis B/sangre , Hepatitis B/diagnóstico , Hepatitis B/virología , Antígenos de Superficie de la Hepatitis B/clasificación , Antígenos de Superficie de la Hepatitis B/genética , Virus de la Hepatitis B/clasificación , Virus de la Hepatitis B/genética , Virus de la Hepatitis B/inmunología , Humanos , Datos de Secuencia Molecular , Juego de Reactivos para Diagnóstico , Sensibilidad y Especificidad , Alineación de Secuencia , Homología de Secuencia de Aminoácido , América del Sur/epidemiología , Estados Unidos/epidemiologíaRESUMEN
OBJECTIVE: To study the analytical sensitivity on 31 HBsAg enzyme immunoassy (EIA) test kits. METHODS: Thirty one HBsAg EIA kits produced by domestic or overseas manufactories and applied for approval during May 2007 to May 2008, were evaluated using the national reference panels. The hyperbolic curve of the log A value and log concentration for the national sensitivity standards was established. The cut-off value of each kit was substituted into the curvilinear equation to determine the analytical sensitivity which was compared between different HBsAg EIA kits. RESULTS: Twenty seven (351 lots) domestic and 4 (27 lots) overseas kits were compared. Among 378 lots of the 31 HBsAg EIA kits, only 2 lots of the domestic kits had a lower sensitivity when tested with the national HBsAg reference panels, with an average approvalr ate of 99.43% (349/351). The mean analytical sensitivity of the domestic kits for adr, adw, ay serotypes were 0.307, 0.419, 0.513 ng/ml, respectively. There was a significant difference between serotypes (F = 97.30, P < 0.01). The mean analytical sensitivity of the overseas kits for adr, adw, ay serotypes were 0.054, 0.066, 0.050 ng/ml respectively, with no significant difference between serotypes (F = 0.65, P > 0.05). The analytical sensitivity of the overseas kits for all the three serotypes was higher than that of the domestic kits (P < 0.01). There was no significant difference found between the analytical sensitivities of the kits produced by the same manufactory using 30- or 60-minute incubation of detection (P > 0.05). In contrast, there was significant difference noticed between the analytical sensitivities of the kits produced by the same manufactory when tested for 10 or 15-minute coloration of the results (P < 0.01). CONCLUSION: Analytical sensitivity of the HBsAg EIA domestic kits should be further improved, especially for detecting adw and ay serotypes.
Asunto(s)
Antígenos de Superficie de la Hepatitis B/aislamiento & purificación , Técnicas para Inmunoenzimas/métodos , Juego de Reactivos para Diagnóstico/normas , Antígenos de Superficie de la Hepatitis B/clasificación , Humanos , Valores de Referencia , Sensibilidad y Especificidad , SerotipificaciónRESUMEN
PreS domain of Hepatitis B virus (HBV) surface antigen is a good candidate for an effective vaccine as it activates both B and T cells besides binding to hepatocytes. This report deals with overexpression and purification of adr subtype of surface antigen that is more prevalent in Pakistan. PreS region, comprising 119 aa preS1 region plus a 55 aa preS2 region plus 11 aa from the N-terminal S region, was inserted in pET21a+ vector, cloned in E. coli DH5alpha cells and expressed in E. coli BL21 codon+ cells. The conditions for over expression were optimized using different concentrations of IPTG (0.01-5 mM), and incubating the cells at different temperatures (23-41 degrees C) for different durations (0-6 h). The cells were grown under the given optimized conditions (0.5 mM IPTG concentration at 37 degrees C for 4 h), lysed by sonication and the protein was purified by ion exchange chromatography. On the average, 24.5 mg of recombinant protein was purified per liter of culture. The purified protein was later lyophilized and stored at -80 degrees.
Asunto(s)
Antígenos de Superficie de la Hepatitis B/genética , Antígenos de Superficie de la Hepatitis B/aislamiento & purificación , Virus de la Hepatitis B/genética , Virus de la Hepatitis B/inmunología , Precursores de Proteínas/genética , Precursores de Proteínas/aislamiento & purificación , Secuencia de Bases , Cartilla de ADN/genética , ADN Viral/genética , Escherichia coli/genética , Expresión Génica , Genes Virales , Inestabilidad Genómica , Antígenos de Superficie de la Hepatitis B/clasificación , Vacunas contra Hepatitis B/genética , Vacunas contra Hepatitis B/aislamiento & purificación , Datos de Secuencia Molecular , Pakistán , Plásmidos/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/aislamiento & purificaciónRESUMEN
Hepatitis B virus (HBV) has been classified into eight genotypes (some of them further divided into two or more subgenotypes) and nine HBsAg subtypes, distinctly distributed geographically. The aim of this study was to gain insight into the distribution of HBV genotypes, subgenotypes and HBsAg subtypes among HBV chronically infected patients in Serbia, since there were no previously published data on this subject. Eighty-nine plasma samples that gave a positive result in a nested PCR were included for genotype identification. Genotyping was performed by direct sequencing of the part of the S/pol gene, and the HBsAg subtype was deduced from the HBsAg sequence. Two HBV genotypes, A and D, were encountered in Serbia, with genotype D (D - 82%, A - 18%) and subgenotype D3 (47.9%) being prevalent. Genotype D isolates had three assigned subtypes (ayw2, ayw3, ayw4), with ayw2 found to be the most prevalent (ayw2 - 53.4%, ayw3 - 43.8%, ayw4 - 1.4%). Genotype A isolates belonged to the A2 subgenotype and the HBsAg subtype adw2, as expected for samples from European population. The results correspond to country's geographical position, being in close proximity to the Mediterranean basin and on the main route between the Middle East and Central Europe.
Asunto(s)
Antígenos de Superficie de la Hepatitis B/clasificación , Virus de la Hepatitis B/clasificación , Virus de la Hepatitis B/genética , Hepatitis B Crónica/epidemiología , Adolescente , Adulto , Anciano , Femenino , Genotipo , Antígenos de Superficie de la Hepatitis B/sangre , Antígenos de Superficie de la Hepatitis B/genética , Virus de la Hepatitis B/aislamiento & purificación , Hepatitis B Crónica/virología , Humanos , Masculino , Persona de Mediana Edad , Datos de Secuencia Molecular , Filogenia , Análisis de Secuencia de ADN , Yugoslavia/epidemiologíaRESUMEN
A seroepidemiological study of the spread of hepatitis B and C viruses (HBV and HCV) was conducted among some population groups in Kabardino-Balkaria. The structure of HBsAg subtypes was also studied in the residents of the republic. The presence of viral hepatitis B markers among the test groups of the healthy population corresponds to the parameters of moderate activity of an epidemic process. The analysis of the immunological structure of the population leads the author to assign Kabardino-Balkaria to highly HBC endemic areas. The occupational factor is demonstrated to be actively involved in the spread of HBC and it is practically of no significance for HCV. An examination could reveal the HBsAg subtype adrq+ that is uncharacteristic of this area.
Asunto(s)
Anticuerpos contra la Hepatitis B/sangre , Antígenos de Superficie de la Hepatitis B/clasificación , Hepatitis B/epidemiología , Anticuerpos contra la Hepatitis C/sangre , Hepatitis C/epidemiología , Adolescente , Adulto , Niño , Preescolar , Antígenos de Superficie de la Hepatitis B/sangre , Humanos , Lactante , Cuerpo Médico , Persona de Mediana Edad , Exposición Profesional , Factores de Riesgo , Federación de Rusia/epidemiología , Estudios SeroepidemiológicosRESUMEN
Hepatitis B virus (HBV) is a human DNA virus, which replicates through an RNA intermediate because of the reverse-transcriptase (RT) activity of its DNA polymerase. As a result, the mutation rate for HBV is higher than the rate observed for most DNA viruses. HBVs are classified into genotypes based on genomic sequencing, and antigenic subtypes based on the antigenic properties of its major surface glycoprotein, the HBV surface antigen (HBsAg). Subgenotypes have been identified within most of the HBV genotypes. The HBV groups defined by the different genotype-HBsAg subtype associations found over the world display characteristic geographical distributions, reflecting the movements of human populations and other epidemiologically significant events. Such HBV groups constitute genetically stable viral populations sharing a common evolutionary history, but additional stable changes, originating from mutation and mutant selection, are observed within all of them. These viral sub-populations are known as the HBV variants, and some of which have medical and public health relevance. Pre-core (pre-C) defective variants have been shown to make HBV infection much less susceptible to interferon treatment, and treatment failures with other antiviral drugs have been associated with selection of resistant variants that display specific mutations in the genome region encoding the viral RT activity. Since the RT region of the genome overlaps the sequence encoding the HBsAg molecule, selection of drug resistant variants involves, in some cases, the indirect selection of HBsAg variants. Viral variants displaying changes in HBsAg seem to be very common among chronic HBV carriers; and some of these variants may emerge under the pressure of the neutralizing antibody response, leading to vaccine resistance and resistance to immunotherapy. Mutations conferring resistance to immunotherapy are noted often among liver transplant recipients and among babies born to HBV-carrier mothers. In addition, some of these HBsAg variants have been associated with lack of detection by HBsAg tests used for the diagnosis of HBV infection, for the identification of chronic carriers, for screening of blood donations for transfusion, and in the manufacture of therapeutic blood products.
Asunto(s)
Virus de la Hepatitis B/genética , Hepatitis B/virología , Antivirales/farmacología , Portador Sano/virología , Farmacorresistencia Viral , Evolución Molecular , Variación Genética , Genoma Viral , Hepatitis B/diagnóstico , Hepatitis B/tratamiento farmacológico , Antígenos de Superficie de la Hepatitis B/clasificación , Antígenos de Superficie de la Hepatitis B/genética , Virus de la Hepatitis B/clasificación , Virus de la Hepatitis B/efectos de los fármacos , Hepatitis B Crónica/virología , Humanos , Trasplante de Hígado/efectos adversos , Mutación , ADN Polimerasa Dirigida por ARN/genética , ADN Polimerasa Dirigida por ARN/metabolismoRESUMEN
The design of a new HBsAg screening assay, the Hepanostika HBsAg Ultra is based on the use of monoclonal antibodies raised against native wild-type HBsAg and reactive with HBsAg in which the common 'a'-determinant is modified by site-directed mutagenesis of four of the cysteine moieties. The design was checked using the same cysteine variants and samples from patients known to be infected with HBsAg variants. The results found were compared with other state-of-the-art commercial screening assays. The design of the Hepanostika HBsAg Ultra enabled detection of all variant HBsAg-positive samples in contrast to the other commercial assays. An additional 980 samples were tested to assess the specificity and sensitivity of the Hepanostika HBsAg Ultra. Screening of presumed negative serum and plasma samples resulted in a specificity of 100%. This makes the Hepanostika HBsAg Ultra the first screening assay with a design able to detect HBsAg variants with high sensitivity and specificity.
Asunto(s)
Ensayo de Inmunoadsorción Enzimática , Antígenos de Superficie de la Hepatitis B/sangre , Hepatitis B/diagnóstico , Animales , Anticuerpos Monoclonales/inmunología , Variación Antigénica/inmunología , Donantes de Sangre , Antígenos de Superficie de la Hepatitis B/clasificación , Antígenos de Superficie de la Hepatitis B/inmunología , Humanos , Ratones , Sensibilidad y EspecificidadRESUMEN
HBs antigens were subtyped in blood samples of intravenous drug-addicts (IDA) and of donors from as many as 10 cities of Western Siberia by using the immune-enzyme assay with 6 high-specific monoclonal antibodies. Two HBsAg subtypes were found, in a ratio of 3% ayw2: 97% ayw3 (varA and varB), in IDA blood samples from Novosobirsk. Three HBsAg subtypes were found, in a ratio of 57% ayw2: 42% ayw3 (varA and varB): 1% adw2, in the donors' blood samples. The obtained data are sufficient for developing the first national sera panel containing different HBsAg subtypes of hepatitis B virus typical of Russia.
Asunto(s)
Antígenos de Superficie de la Hepatitis B/aislamiento & purificación , Antígenos de Superficie de la Hepatitis B/clasificación , Humanos , Técnicas para Inmunoenzimas , Siberia , Abuso de Sustancias por Vía Intravenosa/inmunologíaRESUMEN
A set of monoclonal antibodies with specificity for hepatitis B surface antigen (HBsAg) was used for subtyping this antigen in sera from indigenous natives, blood donors, and drug users in Western Siberia with a modified commercial enzyme immunoassay kit for HBsAg detection. Three subtypes of HBsAg in a ratio of 36 (78%) ayw2:8 ayw3varB (18%):2 (4%) adw2 were found in 46 (100%) HBsAg-positive sera of different aboriginal populations of Western Siberia: the Tundra Nenets, Northern Khanty, Southern Altaians, and Kazakhs. Four subtypes of HBsAg in a ratio of 81 (57%) ayw2:58 (15 ayw3varA and 43 ayw3varB; 44%):2 (1%) adw2 were detected in 141 (100%) samples of blood donors from ten cities of Western Siberia. Three subtypes of HBsAg in a ratio of 34 ayw3:(both variants, 33 ayw3varA and 1 ayw3varB; 97.1%):1 (2.9%) ayw2 were found in blood of 35 injection drug users in Novosibirsk.
Asunto(s)
Antígenos de Superficie de la Hepatitis B/sangre , Antígenos de Superficie de la Hepatitis B/clasificación , Hepatitis B Crónica/epidemiología , Anticuerpos Monoclonales/inmunología , Especificidad de Anticuerpos , Donantes de Sangre , Anticuerpos contra la Hepatitis B/inmunología , Antígenos de Superficie de la Hepatitis B/inmunología , Virus de la Hepatitis B/clasificación , Virus de la Hepatitis B/inmunología , Hepatitis B Crónica/inmunología , Hepatitis B Crónica/virología , Humanos , Técnicas para Inmunoenzimas , Prevalencia , Juego de Reactivos para Diagnóstico , Siberia/epidemiología , Abuso de Sustancias por Vía Intravenosa/complicacionesAsunto(s)
Antígenos de Superficie de la Hepatitis B/genética , Virus de la Hepatitis B/genética , Mutación , Epítopos/genética , Frecuencia de los Genes , Antígenos de Superficie de la Hepatitis B/clasificación , Antígenos de Superficie de la Hepatitis B/aislamiento & purificación , Vacunas contra Hepatitis B/farmacología , Virus de la Hepatitis B/aislamiento & purificación , Humanos , Lactante , SingapurRESUMEN
We described a Hepatitis B surface antigen (HBsAg) subtyping method based on a commercial enzyme immunoassay (EIA) for detection of HBsAg in which the procedure was modified to include the use of monoclonal antibodies with restricted anti-HBs specificities. This method, which was able to classify HBsAg as: ayw1, ayw2, ayw3, ayw3* (intermediate between ayw3 and ayw4), ayw4, ayr, adw2, adw4 and adr, was compared to counter electrophoresis procedure (CEP) by testing HBsAg positive sera from blood donors included in a prospective national epidemiological survey. Among the 256 HBsAg positive samples tested with both techniques, 111 (43.3%) could not be subtyped with CEP vs 10 (3.9%) with our modified EIA. This difference was related to the serum HBsAg concentration which must be greater than 3000 ng/mL and 100 ng/mL for CEP and EIA, respectively. The results obtained from 145 sera with both methods were concordant. Seventeen out of 18 samples partially classified as ay with CEP were completely determined with EIA. This reliable procedure, derived from commercially available reagents, can be easily used in several applications such as large epidemiologic studies and as a substitute for nucleotide sequencing genotyping which is not adapted for large-scale screening and not applicable on samples from nonviremic hepatitis B virus (HBV) carriers.