Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 20
Filter
1.
Article in English | WPRIM | ID: wpr-719646

ABSTRACT

BACKGROUND: We examined changes in hepatitis B core-related antigen (HBcrAg) during the four sequential phases of chronic hepatitis B virus (HBV) infection: hepatitis B e antigen (HBeAg)-positive chronic infection (EPCI) and hepatitis (EPCH), followed by HBeAg-negative chronic infection (ENCI) and hepatitis (ENCH). We compared the performance of serum HBcrAg, hepatitis B surface antigen (HBsAg), and HBV DNA in predicting EPCH and ENCH. METHODS: We enrolled 492 consecutive patients: 49 with EPCI, 243 with EPCH, 101 with ENCI, and 99 with ENCH. HBcrAg was detected by chemiluminescent enzyme immunoassays. HBsAg and HBeAg were detected by chemiluminescent microparticle immunoassays. HBV DNA was detected by real-time PCR. Predictive performance of HBcrAg, HBsAg, and HBV DNA was evaluated using ROC curves. RESULTS: Areas under ROC curves (AUCs) of HBcrAg, HBsAg, and HBV DNA for predicting EPCH were 0.738, 0.812, and 0.717, respectively; optimal cutoffs were ≤1.43×105 kU/mL, ≤1.89×104 IU/mL, and ≤3.97×107 IU/mL, with sensitivities and specificities of 66.3% and 77.6%, 65.0% and 93.9%, and 60.5% and 79.6%, respectively. AUCs of HBcrAg, HBsAg, and HBV DNA for predicting ENCH were 0.887, 0.581, and 0.978, respectively; optimal cutoffs were >26.8 kU/mL, >2.29×102 IU/mL, and >8.75×103 IU/mL, with sensitivities and specificities of 72.7% and 95.1%, 86.9% and 39.6%, and 89.9% and 92.1%, respectively. CONCLUSIONS: HBsAg and HBV DNA were the best predictors of EPCH and ENCH, respectively. HBcrAg is an important surrogate marker for predicting EPCH and ENCH.


Subject(s)
Area Under Curve , Biomarkers , DNA , Hepatitis B e Antigens , Hepatitis B Surface Antigens , Hepatitis B virus , Hepatitis B , Hepatitis B, Chronic , Hepatitis , Hepatitis, Chronic , Humans , Immunoassay , Immunoenzyme Techniques , Real-Time Polymerase Chain Reaction , ROC Curve
2.
Article in Chinese | WPRIM | ID: wpr-491327

ABSTRACT

Objective To investigate the relationship between myeloid-derived suppressor cell ( MDSC) levels and immune function and hepatitis B virus DNA ( HBV-DNA) replication level in patients with hepatitis B. Methods 96 patients with hepatitis B were included in the study.40 normal persons who received physical examina-tion in the hospital during the same period were included as the control group.The content of MDSC was detected by flow cytometry and HBV-DNA level was detected by real-time fluorescence quantitative PCR technique.Statistical analysis was performed.Results The content of MDCS in hepatitis B group (8.77 ±3.69)%was significantly high-er than (1.64 ±1.26)%in the control group (t=16.80,P107 IU/mL ( 90.91%) was significantly higher than that in HBsAg negative group (2.70%)(P<0.05).The content of MDSC in HBV DNA positive group[(10.82 ±3.26)%] was significantly higher than that in HBV DNA negative group[(5.84 ±2.16)%](t=8.42,P<0.05).Spearman's correlation analy-sis showed that the level of MDCS expression was positively correlated with the level of DNA replication (r=0.769, P<0.05).Conclusion The increased level of MDSC content can cause the immune suppression and is positively correlated with the degree of HBV DNA replication.

3.
Article in Chinese | WPRIM | ID: wpr-493705

ABSTRACT

Objective To develop SYBR Green I real-time PCR assay for detection and identification of Hepatitis B virus. Methods Based on the sequences of Hepatitis B virus gp1 gene,primers were designed.The reaction assay and thermal cyc-ling profile were optimized.The positive standard was from recombinant clone.Both the developed assay and Zhejiang kuake biotechnology company’s assay were applied in 100 patients serum.Results The detection limit was between 5×102 copies/ml to 5×108 copies/ml with a good liner correlation and no cross reaction.The whole process just needed 2.5 h.Comparing with the company products,the sensitivity and specificity of the developed assay were 100% and 92.5% respectively.Con-clusion The established assay is rapid,simple,high sensitivity and specificity.It is not only valuable for the identification of Hepatitis B virus patients,but also provide accurate quantitative analysis for HBV patients.

4.
Indian J Med Microbiol ; 2015 Jan-Mar ; 33 (1): 96-100
Article in English | IMSEAR | ID: sea-156997

ABSTRACT

Purpose: Hepatitis B surface Antigen (HBsAg) is the hallmark in diagnosing hepatitis B virus (HBV) infection. In India many commercial assays are available for detection of HBsAg but very few can measure it quantitatively. The present study presents the comparative evaluation of two methods and their correlation with serum HBsAg in chronic hepatitis B (CHB) patients. Materials and Methods: Consecutive patients of CHB were included and there HBsAg levels were measured by two methods: (i) Elecsys, Roche Diagnostics, a qualitative assay and (ii) Architect, Abbott Diagnostics, a quantitative assay. The HBV DNA was measured by real-time polymerase chain reaction (qPCR). Results: Total of 136 patients were included in the study and there was a signifi cant overall correlation between both the assays (correlation coeffi cient [r] = 0.83; P < 0.001). Assays correlated well with each other across all subgroups of CHB: treatment naïve (r = 0.73; P < 0.001, n = 32), on treatment (r = 0.56; P < 0.05, n = 104), hepatitis Be (HBe) antigen positive (r = 0.67; P < 0.001, n = 62) and anti-HBe positive (r = 0.61; P < 0.05, n = 74) group. On correlation with serum HBV DNA, Architect assay demonstrated good correlation (r = 0.73; P < 0.001, n = 136) as compared to the Elecsys assay (r = 0.27; P = 0.068, n = 136). Architect HBsAg QT assay (A1) also correlated well with HBV DNA in the treatment naïve group (r = 0.69; P < 0.001, n = 32). Conclusions: Our study hence proved that both the assays are comparable and a simple qualitative assay with in-house modifi cation can be used easily for quatitation of HBsAg in clinical samples.

5.
Article in Chinese | WPRIM | ID: wpr-476079

ABSTRACT

Objective To investigate the risk factors of intrauterine hepatitis B virus (HBV)infection and its preventive measures.Methods 254 cases of pregnant women with positive HBsAg and their neonatuses from Shaanxi People’s Hospi-tal were selected as the research object.The serum of pregnant women were tested HBV markers and HBV-DNA,their neo-natal umbilical cord blood were only detected HBV markers.Results In 254 samples,the positive rates of HBsAg and HBeAg within their neonatal umbilical cord blood were 5.12% and 13.78% respectively;the positive rate of HBV infection in neonatal umbilical cord blood among maternal HBeAg-positive group was 53.33%,far higher than 3.61% in HBeAg-neg-ative group (χ2 =99.44,P <0.001);the positive rates of HBsAg and HBeAg in neonatal umbilical cord blood were elevated along with the increase of maternal HBV-DNA copy (Hc= 13.50,66.70;P < 0.001).The positive rate of HBV-DNA in pregnant women with HBeAg-positive group was 100%,far more than 19.59% in HBeAg-negative group (P <0.001).Con-clusion Both maternal positive HBeAg and HBV-DNA are risk factors for neonatal intrauterine HBV infection.Prenatal HBeAg screening is predominant,especially in some hospitals without real-time quantitative assay.An effective measure to reduce intrauterine HBV infection is to be pregnant when HBV DNA and HBeAg are turned into negative after HBV inter-vention and treatment for women in reproductive age.

6.
Article in Chinese | WPRIM | ID: wpr-603227

ABSTRACT

Objective To explore the effect of preoperative antiviral therapy on hepatitis B virus ( HBV ) reactivation and postoperative liver function in perioperative patients with HBV-DNA-negative hepatocellular carcinoma(HCC).Methods 74 patients with preoperative HBV-DNA-negative scheduled which were analyzed.Patients were divided into two groups according to antiviral therapy or not:20 cases in antiviral treatment group received antiviral therapy for three days, 54 cases in non-antiviral teatment group did not receive antiviral therapy, and both groups received antiviral therapy after post-operative resuming to diets.The indicators of liver function and HBV-DNA levels were detected on pre-operative, post-operative 3rd and 7th day in two groups, and HBV-DNA-positive ( HBV-DNA>500 IU/mL) was defined as reactivation, conversely as inactivation.The indicators of liver function on pre-operative, post-operative 3rd and 7th day were compared between reactivation group and inactivation group.Results The reactivative rate was 21.6%(16/74) in all patients;27.7%(15/54) in pre-operative non-antiviral teatment group, 5.0%(1/20) in antiviral teatment group, and there was significant differences in reactivative rate between two groups ( P=0.035 ).The results of Logistic regression showed that pre-operative nonantiviral therapy was an independent risk factor of post-operative HBV reactivation (OR=13.952,95% confidence interval[CI]:1.358-143.379,P=0.027).The recovery of albumin (ALB) on post-operative 3rd, 7th days in antiviral treatment group was faster than those in nonantiviral treatment group, respectively (P=0.035,0.043).The recovery of ALB and alanine aminotransferase (ALT) on post-operative 7th day in reactivation group were slower than those in inactivation group, respectively (P=0.016, 0.048).Conclusion The pre-operative nonantiviral therapy is an independent risk factor of post-operative HBV reactivation in patients with HBV-DNA-negative HCC.The pre-operative antiviral therapy could inhibit post-operative HBV reactivation effectively and accelerate the post-operative recovery of liver function.

7.
Journal of Medical Research ; (12): 88-90,41, 2015.
Article in Chinese | WPRIM | ID: wpr-601881

ABSTRACT

Objective In this research with the method of Alu -PCR we investigate the integration of hepatitis B virus ( HBV) DNA in human hepatocarcinoma cell line (PLC/PRF/5) genome DNA.Methods We at first extracted the genome DNA from PLC/PRF/5 cells, and then the potential integration fragments of HBV DNA and human genome DNA were amplified with according to the Alu -PCR after three rounds PCR .The Alu-PCR amplification products were observated with agarose gel electrophoresis , then integration positive electrophoresis bandings were chipped and purified for nucleic acid sequencing .At last he bioinformatics information was acquired by blast online.Results Through agarose gel electrophoresis after Alu -PCR amplification, we got four potential integration bindings , among which we got three integration sequences of HBV DNA in human genome DNA .These integration sequences could be individually located in the human chromosome of 03p21.31, 05p15.33, 12q13 and 12-q14.1.Conclusion With Alu-PCR we can accurately measure the integration of HBV DNA in human genome DNA , and Alu-PCR can be a a convenience and economic method in the study of HBV DNA ′s integration in human genome DNA .

8.
Rev. cuba. med. trop ; 64(3): 290-303, jul.-sep. 2012.
Article in Spanish | LILACS | ID: lil-653847

ABSTRACT

Introducción: los niveles de ADN viral en muestras de suero son un marcador útil para monitorear la progresión de la enfermedad y la respuesta al tratamiento en pacientes con hepatitis B crónica; de ahí que se comercialicen estuches diagnósticos para esta función, con la desventaja de ser costosos. Objetivos: desarrollar y evaluar el desempeño analítico de un sistema de reacción en cadena de la polimerasa en tiempo real para la cuantificación del ADN del virus de la hepatitis B. Métodos: se utilizaron cebadores que amplifican un fragmento del gen C y sonda de hidrólisis en el equipo LightCycler 1.5. Se construyó una curva estándar y se evaluó su eficiencia. Se utilizaron 272 muestras de suero para ensayos de especificidad analítica y clínica, especificidad y exactitud genotípica, coeficientes de variación intraensayo e interensayo, comparación con un estuche comercial y con la reacción en cadena de la polimerasa cualitativa para el virus de la hepatitis B. Resultados: la curva estándar mostró excelente correlación lineal (r= -1) y valores muy bajos de error a lo largo de varias magnitudes de concentración de ADN diana. La especificidad analítica y clínica fue de 100 %, en tanto que al evaluar la especificidad y exactitud genotípica, se obtuvo que las diferencias entre los Log10 del valor obtenido y el de referencia eran inferiores a 0,5 Log10. El límite de detección por análisis de Probit se estimó en 16,41 UI/µL con un rango dinámico de cuantificación de hasta 10(8) UI/mL. El sistema mostró bajos coeficientes de variación intraensayo (0,16 a 1,45 %) e interensayo (0,9 a 2,62 %). La comparación con el estuche comercial artus HBV LC PCR kit mostró una correlación de r= 0,964 y r²= 0,929; con la reacción en cadena de la polimerasa cualitativa se confirmó la mayor sensibilidad y ventajas de la reacción en cadena de la polimerasa en tiempo real. Conclusiones: el ensayo cumple con los requisitos para la cuantificación del ADN del virus de la hepatitis B, que demuestra ser específico, sensible y reproducible. Su aplicación permitirá un mejor diagnóstico y seguimiento de los pacientes con hepatitis B crónica en Cuba.


Introduction: viral DNA levels in serum samples are a useful marker to monitor the disease progression and the treatment response in patients with chronic hepatitis B. Commercial kits for this purpose are available, but they are considerably expensive. Objectives: to evaluate the analytical performance of a real-time polymerase chain reaction (RT-PCR) assay for Hepatitis B virus DNA quantification. Methods: specific primers to the gene C and TaqMan chemistry in a LightCycler 1.5 equipment was used. A standard curve was made and evaluated. Two hundred and seventy-two serum samples were used to assess the clinical and analytical specificity, the genotypic accuracy and specificity, the intra-assay and interassay coefficients of variation and the comparison with a commercial assay and with the qualitative PCR. Results: the standard curve showed a strong linear correlation (r= -1) and low error values in the tested target DNA concentration. Analytical and clinical specificities were 100 %. Genotype accuracy and specificity showed that the differences between the results obtained by RT-PCR assay and those of the reference assay were less than 0.5 Log10. The 95% HBV DNA detection end-point assessed by Probit analysis was 16.41 IU/µL with a dynamic range of quantification of 10(8) IU/mL. Intra-assay and interassay coefficients of variation ranged from 0.16 to 1.45 % and 0.9 to 2.62 % respectively. The RT-PCR assay correlated well with those from a commercial assay (r= 0.964 and r²= 0.929) and with the HBV qualitative PCR, thus confirming its better sensitivity and advantages. Conclusions: the RT-PCR assay is well suited to monitoring HBV DNA levels showing to be sensitive, specific and reproducible. Its application in the clinical practice ensures a better diagnosis and management of patients with chronic hepatitis B in Cuba.


Subject(s)
Humans , DNA, Viral/analysis , Hepatitis B virus/genetics , Real-Time Polymerase Chain Reaction
9.
Indian J Med Microbiol ; 2012 Apr-June; 30(2): 150-154
Article in English | IMSEAR | ID: sea-143936

ABSTRACT

Purpose : The hallmark of chronic hepatitis B (CHB) infection is the presence of hepatitis B surface antigen (HBsAg) positivity for at least 6 months. Recently, serum levels of HBsAg have been compared with serum HBV DNA as a surrogate marker to monitor CHB patients. However, data correlating these two markers are scarce. Hence, the present study was done to correlate HBV DNA with HBsAg in CHB patients. Materials and Methods: Consecutive patients of CHB were included. HBV DNA was measured by real-time polymerase chain reaction (PCR). Serum HBsAg was measured by Architect HBsAg. Results: Of the 198 patients enrolled, 166 fulfilled the inclusion criteria (mean age 43 ± 14 years, 87% males) and the median HBV DNA was 1.7 × 10 3 (range 6.0-1.1 × 10 8 ) IU/ml. Median HBsAg was 8.7 × 10 3 (range 5.0-3.2 × 10 5) IU/ml. Overall correlation between HBV DNA and HBsAg was weak but significant (Spearman ρ = 0.443, P < 0.01). Correlation in HBe antigen-positive group was better (ρ = 0.402, P < 0.01) in comparison to HBe antigen-negative group (ρ = 0.193 P = 0.05). Good correlation existed in treatment-naïve group (ρ = 0.538, P < 0.01) .Correlation was regardless of normal or raised alanine transaminase (ALT). Eighty (48%) patients had high HBV DNA (≥2000 IU/ml). Correlation in high DNA group was significant (P < 0.01). The best cut-off of HBsAg for diagnosing high DNA is 3.36 ×10 3 IU/ml. Conclusions: Serum HBsAg correlates with HBV DNA in CHB patients, especially in high serum HBV DNA, HBe antigen-positive and treatment-naïve group. HBsAg levels can be used for predicting high serum HBV DNA levels.


Subject(s)
Adolescent , Adult , Aged , Child , Child, Preschool , Cross-Sectional Studies , DNA, Viral/blood , Hepatitis B Surface Antigens/blood , Humans , Immunoassay , Male , Middle Aged , Prospective Studies , Real-Time Polymerase Chain Reaction , Serum/chemistry , Statistics as Topic , Young Adult
10.
Korean Journal of Medicine ; : 731-739, 2012.
Article in Korean | WPRIM | ID: wpr-211750

ABSTRACT

BACKGROUND/AIMS: The prevalence of occult HBV infection (OBI) in patients with chronic hepatitis C (CHC) in Korea has not been reported. Additionally, it is unclear whether OBI influences treatment outcome in CHC patients. We investigated the prevalence of OBI and its impact on treatment outcome in patients with CHC. METHODS: Seventy-six patients with CHC were enrolled and treated with pegylated or conventional interferon and ribavirin. Hepatitis B virus (HBV) DNA was detected by nested polymerase chain reaction. RESULTS: Among the 68 patients who completed treatment and follow-up, HBV DNA was detected in serum from nine (13.2%) patients, liver tissue from 10 (14.7%), and serum or liver tissue from 15 (22.1%). OBI was diagnosed in nine (12.7%) control subjects. No difference in the prevalence of OBI between patients with CHC and controls was observed (13.2 vs. 12.0%; p = 0.92). No significant differences in age, sex, genotype 1 frequency, amount of hepatitis C virus RNA, anti-hepatitis B surface antigen/anti-hepatitis B core-IgG seropositivity, staging, or histology grading were observed in patients with or without HBV DNA. Sustained virological response was achieved in 73.3% of patients with OBI and 83.0% without OBI (p = 0.46). CONCLUSIONS: These results demonstrate that a significant proportion of patients with CHC have occult HBV infection and that OBI does not affect treatment outcome in patients with CHC.


Subject(s)
DNA , Follow-Up Studies , Genotype , Hepacivirus , Hepatitis B virus , Hepatitis C, Chronic , Hepatitis, Chronic , Humans , Interferons , Korea , Liver , Prevalence , Ribavirin , RNA , Treatment Outcome
11.
Article in Korean | WPRIM | ID: wpr-151920

ABSTRACT

Over the past decade, advances in the antiviral therapy in patients with chronic hepatitis B have enabled the sustained suppression of hepatitis B viral replication and the prevention of progressive liver disease. Hepatitis B surface antigen (HBsAg) has been used to diagnose patients with hepatitis B virus infection. Recently, test for quantitative HBsAg titers are available and on-treatment HBsAg quantitations are used to predict treatment outcome. Serum HBV DNA levels have been shown to predict natural course of chronic hepatitis B infection. The HBV DNA levels have been reported to be positively correlated with the development of cirrhosis, hepatocellular carcinoma and related death. The baseline and on-treatment levels of HBV DNA are important factors for predicting treatment outcomes. In this article, we will discuss the role of HBV DNA and HBsAg quantitation during antiviral therapy.


Subject(s)
Antiviral Agents/therapeutic use , Carcinoma, Hepatocellular/etiology , DNA, Viral/blood , Hepatitis B Surface Antigens/blood , Hepatitis B virus/genetics , Hepatitis B, Chronic/complications , Humans , Lamivudine/therapeutic use , Liver Cirrhosis/etiology , Liver Neoplasms/etiology , Nucleosides/therapeutic use , Pyrimidinones/therapeutic use
12.
Chinese Journal of Epidemiology ; (12): 159-162, 2010.
Article in Chinese | WPRIM | ID: wpr-277668

ABSTRACT

Objective To study the influence of HBV-DNA with different load levels of HBsAg-positive among fathers on the rate of neonatal cord blood HBV-DNA.Methods Using HBsAg and HBV-DNA as screening indicators for pregnant women and their husbands from an obstetric clinic.161 pregnant women whose HBsAg and HBV-DNA were negative,but HBsAg was positive among their husbands and their newborns,were selected.Blood samples from those pregnant women,their husbands and their newborns were collected to detect the related indicators.Using ELISA to detect hepatitis B virus markers(HBVM),and FQ-PCR to detect the levels of HBV-DNA load.According to neonatal cord blood HBV-DNA detection guideline,newborns with cord blood HBV-DNA positive were selected as cases,others as controls.Results(1)Result of the study showed that there was a dose-response relationship between paternal serum HBV-DNA load levels and neonatal cord blood HBV-DNA positive rates in newborns(trend χ~2=64.117,P=0.000).The rate of vertical transmission of HBV from HBsAg-positive father to infant in the paternal serum HBV-DNA>1.0×107 copies/ml group was significantly higher than HBV-DNA<1.0×107 copies/ml group(χ~2=71.539,P=0.000).(2)There was a positive rank correlation between semen positive HBeAg and vertical transmission of HBV from HBsAg-positive father to infant(χ~2=6.892,P=0.009).Conclusion There was a dose-response relationship between paternal serum HBV-DNA load levels and neonatal cord blood HBV-DNA positive in newborns.Paternal serum HBV-DNA≥1.0×107 copies/ml and with HBeAg positive status were risk factors of vertical transmission of HBV from HBsAg-positive father to infant.

13.
Rev. Soc. Bras. Med. Trop ; 41(6): 596-601, Nov.-Dec. 2008. ilus, tab
Article in Portuguese | LILACS | ID: lil-502041

ABSTRACT

Recentemente é descrito estado de persistência do vírus da hepatite B denominado hepatite crônica B oculta. Sua prevalência e fisiopatologia são desconhecidas. O objetivo deste estudo foi avaliar a ocorrência dessa entidade clínica em pacientes da Amazônia brasileira. De 51 pacientes anti-HBc total reativos testados pela reação em cadeia da polimerase, 17 por cento foram positivos. Não observamos associação com fatores de risco clássicos de infecção pelo vírus da hepatite B, testes bioquímicos, hematológicos e histopatologia. No entanto, os pacientes ictéricos e reativos para o anti-HIV apresentaram associação com a presença do ADN-vírus da hepatite B. Os resultados demonstram a ocorrência da hepatite crônica B oculta, entre nossos doentes, porém, com taxas de prevalência abaixo do esperado para a região. Acreditamos que, apesar do tamanho da amostra avaliada ser pequeno, sua ocorrência poderia ter sido maior se empregássemos primers para a região S, C e X do genoma do vírus da hepatite B, aumentando a sensibilidade do teste.


A persistent form of the hepatitis B virus called occult chronic hepatitis B has recently been described. Its prevalence and physiopathology are unknown. The aim of this study was to evaluate the occurrence of this clinical entity among patients in the Brazilian Amazon region. Out of 51 anti-HBc total-positive patients who were tested using the polymerase chain reaction, 17 percent were positive. We did not find any associations with classical risk factors for hepatitis B virus infection or with biochemical tests, hematological tests or histological patterns. However, the jaundiced and HIV-positive patients showed a statistical association with the presence of hepatitis B virus-DNA. The results demonstrated that occult hepatitis B occurred among our patients, but at prevalence rates lower than expected for this region. We believe that despite the small sample size, the occurrence might have been found to be greater if we had used primers for the S, C and X regions of the hepatitis B virus genome, thereby increasing the sensitivity of the test.


Subject(s)
Female , Humans , Male , Endemic Diseases , Hepatitis B Antibodies/blood , Hepatitis B Core Antigens/immunology , Hepatitis B virus/isolation & purification , Hepatitis B, Chronic/epidemiology , Brazil/epidemiology , DNA, Viral/analysis , Hepatitis B virus/genetics , Hepatitis B virus/immunology , Hepatitis B, Chronic/diagnosis , Polymerase Chain Reaction , Prevalence , Prospective Studies , Risk Factors
14.
Article in Chinese | WPRIM | ID: wpr-685970

ABSTRACT

Objective:To construct a DNA vaccine co-expressing the HBV compound multi-epitope antigen gene, the hIL-12 and the anti-HBV siRNA genes, and to express this DNA vaccine in HepG2 cells. Methods:The HBV multi-epitope antigen gene was designed and synthesized before it was fused with enhanced green fluorescent protein(EGFP) gene, and cloned into the multi-clone site(MCS) of the eukaryotic expression vector pVAX1. The expressinig units of hIL-12 and siRNA were cloned into the BspH I and Mlu I site of pVAX1 respectively. Then the recombinant plasmid pVAX1-siHBV-HB-EGFP-hIL12 was transiently transfected HepG2 cells. The expression of HBV compound multi-epitope gene was observed through EGFP report gene. The expression of hIL-12 was analyzed by ELISA and the effects of anti-HBV siRNA was confirmed with rtPCR . Results: The analysis of enzyme digestion and sequencing both demonstrated that the trible-expressing HBV DNA vaccine has been constructed successfully. The green fluorescent image was detected in the transfected cells which could confirm the expression of the multi-epitope antigen gene. The amount of hIL-12 secretion was 1289pg/ml in supernatant at 48h after transfection and 1712pg/ml at 72h after transfection. The mRNA amount of HBV S gene, which was the siRNA target, had been obviously knockdown. Conclusion: The DNA vaccine co-expressing the HBV compound multi-epitope antigen gene, the hIL-12 and the siRNA genes was constructed and transiently expressed in HepG2 cells, and siRNA had shown us a good anti-HBV effect. It laid a foundation of further study on anti-HBV effect of the new DNA vaccine.

15.
Article in Chinese | WPRIM | ID: wpr-639170

ABSTRACT

Objective To investigate the efficacy by inoculated with hepatitis B immunoglobulin(HBIG) and hepatitis B vaccine in dif-ferent doses and time points to prevent maternal-infant vertical transmission of hepatitis B virus(HBV).Methods Gravidas positive for HBV were selected and measured HBV DNA,according to difference HBV DNA degrees divided into A,B,C groups.Every group was randomly divided into 4 groups using a variety of combined immunity methods to compare its efficacies.Results There were no significant differences in the positive rate of HBV and HBsAb in A,B groups by statistics analysis.There were significant differences in the positive rate of HBV and HBsAb in group C by statistics analysis.Conclusion According to the different HBV DNA take different combined immunodeficiency approach to prevent maternal-infant vertical transmission.

16.
Article in Chinese | WPRIM | ID: wpr-584056

ABSTRACT

Objective To study suitability of a series of lyophilized serum with definitive HBV DNA value in fluorescence quantitative COBAS Amplicor HBM kit and a sample with 106 copies/ml HBV DNA was prepared, and sent to various manufactures which would be asked to detect the samples using their own kits. Then a calibration curves from CT values of the series to the corresponding concentrations was compared with that obtained from the external standard-calibration curve with the manufactures series. Results The standard-calibration curve with the series of lyophilized serum showed an excellent correlation (

17.
Article in Chinese | WPRIM | ID: wpr-522906

ABSTRACT

0 05). Between the patients with high level of serum HBV-DNA and the ones with low level of serum HBV-DNA, both the quantity and positive rate of HBV-DNA in PBMC had a significant difference (P

18.
Korean Journal of Anatomy ; : 629-635, 1999.
Article in Korean | WPRIM | ID: wpr-647054

ABSTRACT

In the present study author investigated 48 patients with chronic hepatitis B for the presence of peripheral blood HBV-DNA with the aid of DNA molecular techniques. HBV-DNA was detected in peripheral blood cells in 34 of 48 (70.8%) of chronic HBsAg positive patients. In order to know that the presence of HBV-DNA in peripheral blood induce more chromosomal instability in comparison with the absent, frequency of SCE was analyzed in peripheral blood. In HBV-DNA positive chronic hepatitis B patients a mean frequency of SCE was 11.5452+/-0.6944, in HBV-DNA negative patients 11.7540+/-0.7032, respectively. Both groups had significantly higher SCE frequence than that of normal control group (5.8533+/-0.437) (p<0.05). There was no significant difference between them. This study may suggest that the presence of HBV-DNA in peripheral blood of patients with chronic hepatitis B may be related to certain pathogenetic processes in HBV infection, but it may have no effect on chromosomal instability in peripheral blood.


Subject(s)
Blood Cells , Chromosomal Instability , DNA , Hepatitis B Surface Antigens , Hepatitis B virus , Hepatitis B , Hepatitis B, Chronic , Hepatitis , Hepatitis, Chronic , Humans , Siblings , Sister Chromatid Exchange
19.
Article in Korean | WPRIM | ID: wpr-229236

ABSTRACT

BACKGROUND: Quantitative measurement of hepatitis B virus (HBV) DNA in serum allows monitoring of HBV replication and assessing responses to antiviral treatment, which is not possible with serological markers for HBV infection. However, most assays developed for the quantitative measurement of HBV DNA have not been standardized. Therefore, we tried to compare the performance of three commercial quantitative methods for the measurement of HBV DNA. METHODS: One hundred consecutive sera with request for HBV DNA quantitation were tested with Hybrid Capture System (HCS), Hybrid Capture II (HC-II) and Quantiplex HBV DNA Assay (bDNA Assay) to evaluate the detection rate, the concordance rate and the correlation of the quantitative results measured by each method. In addition, nested polymerase chain reaction (PCR) was performed for qualitative detection of HBV DNA. RESULTs: Concordance rate was 87% for all three methods, 91% for HCS and HC-II, 94% for HCS and bDNA Assay, and 89% for HC-II and bDNA Assay. HBV DNA quantities measured by three methods showed significant correlation between HCS and HC-II (R=0.88, P<0.0001), HCS and bDNA Assay (R=0.82, P<0.0001), and HC-II and bDNA Assay (R=0.95, P<0.0001). Thirteen sera of discrepant results and 29 of 39 sera of negative results by all three methods showed PCR positivity. CONCLUSIONS: Three quantitative methods for the measurement of HBV DNA showed relatively high concordance rate and good correlation. However, the results by bDNA Assay increased more rapidly than HCS and HC-II as the amounts of HBV DNA in the sample increased that the concentrations of HBV DNA measured by bDNA Assay were two or three times higher than those measured by HCS or HC-II at high HBV DNA concentration range. Thus, further studies are necessary to develop more standardized methods.


Subject(s)
Branched DNA Signal Amplification Assay , DNA , Hepatitis B virus , Hepatitis B , Hepatitis , Polymerase Chain Reaction
20.
Article in Korean | WPRIM | ID: wpr-55075

ABSTRACT

In order to examine a possibility of hepatitis B virus(HBV) transmission via tears, detection of HBV DNA in tear was esamined using polymerase chain reaction (PCR) in 39 patients with HBsAg positive in serum. The detection of HBV DNA in tears was compared that in serums. The patients were divided into group I (n=21, serum HBsAg+HBeAg-) and group II (n=18, serum HBsAg+/HBeAg+). The total detection rates of HBV DNA in tears and serums were 76.9% and 92.3%, respectively. In detail, detection rates of HBV DNA in tears were 71.4% (group I) and 83.3% (group II) and those in serums were 85.7% (group I)and 100% (group II). There was no statisticlly significant difference in detection rate between group I and group II (p>0.05). However, mean values for serum aminotransferase activities in group II were higher than those in group I (p<0.05). The titer of HBsAg or HBeAg did not influence on the detection rate of HBV DNA. With these results, it was found that HBV DNA would be easily detected in tear of patients with HBV infection and HBV could be transmitted via ophthalmic instruments touched with ocular surface. Therefore, history taking concerning hepatitis B, screening test for hepatitis B, chemical or physical sterilization of ophthalmic instruments, and vaccination for workers in ophthalmologic department would be important in prophylactic aspects.


Subject(s)
Antigens, Surface , DNA , Hepatitis B e Antigens , Hepatitis B Surface Antigens , Hepatitis B virus , Hepatitis B , Hepatitis , Humans , Mass Screening , Polymerase Chain Reaction , Sterilization , Vaccination
SELECTION OF CITATIONS
SEARCH DETAIL