Background: Hepatitis B virus (HBV) infection is one of the worldwide public health problem affecting about 300 million people. The envelope protein of HBV consists of three components known as preS1, preS2, and S antigen. According to the recent study, anti-HBs Ab showed effective neutralization ability against HBV from chronic hepatitis B and liver transplant patients, suggesting the possible development of therapeutic antibody. Methods: Spleen cells immunized with S antigen of HBV were fused with myeloma cell line to obtain HBsAg specific monoclonal antibodies. High affinity antibodies against HBsAg (adr, ad and ay type) were selected by competitive ELISA method. Nucleotide sequence of the variable regions of monoclonal antibodies was analyzed by RT-PCR followed by conventional sequencing method. Results: We produced 14 murine monoclonal antibodies which recognize S antigen of HBV. Two of them, A9-11 and C6-9 showed the highest affinity. The sequence analysis of A9-11 revealed that variable regions of the heavy chain and light chains are members of mouse heavy chain I (B) and light chain lambda 1, respectively. Likewise, the sequence analysis of C6-9 revealed that variable regions of the heavy chain and light chains are members of mouse heavy chain II (B) and light chain kappa 1, respectively. Neutralization assay showed that A9-11 and C6-9 effectively neutralize the HBV infection. Conclusion: These results suggest that A9-11 and C6-9 mouse monoclonal antibodies can be used for the development of therapeutic antibody for HBV infection.
Kim, Soon-Duck;Choi, Ji-Ho;Kim, Sung-Ryul;Lee, Jin-Soo;Koh, Hee-Jeong
Journal of Preventive Medicine and Public Health
/
v.38
no.2
/
pp.170-174
/
2005
Objectives: The aim of this study was to evaluate the response to a hepatitis B vaccination, and investigate the HBV DNA in subjects with isolated anti-HBc. Methods: 34 subjects with persistent isolated anti-HBc were included in the study. 32 subjects negative for HBsAg, anti-HBs and anti-HBc were included as a control group. They were all vaccinated with Hepaccine at 0, 1 and 2 months, and anti-HBs titers were measured 1 month after the 1st and 3rd vaccinations (1 and 3 months). The HBV-DNA was tested by polymerase chain reaction in subjects with isolated anti-HBc. Results: After the 1st & 3rd vaccinations, the anti-HBs titers$\geq$10mIU/ml were 70.6 & 70.6% in isolated anti-HBc group, and 34.4 & 81.2% in the control group, respectively. There were statistically significant differences after the 1st vaccination, but none after the 3rd, between the two groups. In the isolated anti-HBc and control groups, the primary, amnestic and no responses were 0 vs. 46.9%, 55.9 vs. 6.3% and 29.4 vs. 18.8%, respectively. The HBV DNA was not detected in all subjects with isolated anti-HBc. Conclusion: None of the subjects with isolated anti-HBc had a false positive result (primary response); therefore, they should be excluded from vaccination programs in Korea. To differentiate between immunity and occult infections, a single dose of vaccine, with a follow-up anti-HBs test, is preferable for subjects with isolated anti-HBc. An amnestic response indicates late immunity, and no response a suspect occult infection.
In the past, hepatitis B virus (HBV) infection was endemic in the general Korean population. The association of HBV infection with the occurrence of liver cancer has been well demonstrated in several epidemiologic studies. While the mortality rates of liver cancer in Korea have decreased steadily over the last decade, the presence of hepatitis B surface antigen (HBsAg) in mothers remains high at 3-4%, and 25.5% of these HBsAg positive mothers are positive for hepatitis B e antigen (HBeAg). HBV infection caused almost a quarter of hepatocellular carcinoma (HCC) cases and one-third of deaths from HCC. These aspects of HBV infection prompted the Korean government to create a vaccination program against HBV in the early 1980s. In 1995, the Communicable Disease Prevention Act (CDPA) was reformed, and the government increased the number of HBV vaccines in the National Immunization Program (NIP), driving the vaccination rate up to 95%. In 2000, the National Health Insurance Act (NHIA) was enacted, which provided increased resources for the prevention of perinatal HBV infection. Then in 2002, the Korean government, in conjunction with the Korean Medical Association (KMA), launched an HBV perinatal transmission prevention program. The prevalence of HBsAg in children had been high (4-5%) in the early 1980s, but had dropped to below 1% in 1995, and finally reached 0.2% in 2006 after the NIP had been implemented. After the success of the NIP, Korea finally obtained its first certification of achievement from the Western Pacific Regional Office of the World Health Organization (WPRO-WHO) for reaching its goal for HBV control. An age-period-cohort analysis showed a significant reduction in the liver cancer mortality rate in children and adolescents after the NIP had been implemented. In addition to its vaccination efforts, Korea launched the National Cancer Screening Program (NCSP) for 5 leading sites of cancer, including the liver, in 1999. As a consequence of this program, the 5-year liver cancer survival rate increased from 13.2% (1996-2000) to 23.3% (2003-2008). The development of both the primary and secondary prevention for liver cancer including HBV immunization and cancer screening has been of critical importance.
Purpose: This study was undertaken to investigate the feasibility of using automated liquid-handling systems equipped with reusable fixed tips in Radioimmunoassays and to demonstrate that the use of an automated pipetting instrument can decrease in the typical daily workload. Materials and Methods: The precisions of the automated pipettor and the manual pipettor were determined gravimetrically (n=30). A total of 30 specimens for HBs Ag were repeatedly pipetted (three replicates) with the automated pipettor and then retested. PSA samples were simultaneously pipetted with either the automated pipettor or the manual pipettor and then analyzed (n=40). Sample carryover test assessed for CA19-9, AFP and HCG when the automated pipettor was used. Pipetting speed of the automated pipettor and the manual pipettor were compared by evaluation of each workload. Results: The precisions coefficients of variation (CV) were 2.1% for the automated pipettor and 1.6% for manual pipettor. The mean cpm and CV for each group of replicates were 41,203 cpm and 3.7% for HBs Ag positive specimens, and 99 cpm and 7.9% for HBs Ag negative specimens, respectively. PSA results showed no significant differences between automated pipettor and manual pipettor (p=0.15, r=0.999). Carryover for CA19-9, AFP and HCG analytes was <0.1 ppm or below the assay limit of detection. Pipetting speed was significantly improved by using the automated instrument. Conclusion: There was no evidence that the use of an automated pipettor adversely affected any of the performance characteristics of the assay. Indeed, routine use of the Tecan automated pipettor has resulted in a decrease in the typical daily workload.
Lee, Sun-A;Yoo, So Young;Kay, Kee-Sung;Kook, Joong-Ki
Journal of Microbiology
/
v.42
no.3
/
pp.239-242
/
2004
This study examined the detection rate of the hepatitis B virus (HBV) and Mycobacterium tuberculosis (Mtb) in serum and saliva samples, respectively, from 120 dental patients who were unaware if they have or had either hepatitis or tuberculosis. The frequencies of HBsAg and anti-HBs were determined using an immunochromatic assay. Mtb positivity was determined by the PCR method. Of the 120 patients, 7 (5.8%) were HBV positive and 30 (25.0%) were Mtb positive. This highlights the fact that dental health care workers (DHCWs) can be exposed to the risk of infection from blood- or saliva-borne pathogens as a consequence of their work. Therefore, it is very important to prevent cross infection between patients and dental personnel. Accordingly, laboratory tests prior to surgical treatment are needed to determine the infectious state of dental patients in order to prevent the transmission of infectious diseases in dental clinics.
The purpose of this study was to survey the knowledge and awareness of dental hygienists and dental hygiene students about hepatitis B in an attempt to lay the solid foundation for the prevention of hepatitis B and infection control, as dental hygienists were highly likely to be exposed to HBV during job performance. The findings of the study were as follows: 1. 32.5% of the dental hygienists and 15.6% of the dental hygiene students were aware of their own HBsAg and HBsAb states. The dental hygienists who were cognizant of the states statistically significantly outnumbered the dental hygiene students who were (p=.000). 2. As for vaccination against hepatitis B, 71.8% of the dental hygienists and 47.3 % of the dental hygiene students were inoculated against it. The dental hygienists who were vaccinated against it outnumbered the dental hygiene students who were, and the disparity between the two was statistically significant (p=.000). 3. In the event of those who were vaccinated against hepatitis B, the dental hygienists were better cognizant than the dental hygiene students of the right time for the vaccination, required vaccination frequency (three times) and confirmation of the formation of hepatitis B antibody (p=.000). 4. Regarding awareness of hepatitis B infection route, the dental hygienists knew significantly better than the dental hygiene students that hepatitis B might be infected via blood (p=.030), sexual relations (p=.000), contaminated needle sticks (p=.000), mothers with hepatitis B positive during delivery (p=.000), toothbrushes/razors (p=.000) and exchange of drinking cups (p=.000). 5. As to the relationship between health status and knowledge on hepatitis B infection route, the respondents who were in bad shape had the best knowledge about that, followed by those in an average state of health and healthy respondents. And the dental hygienists had a significantly better knowledge than the dental hygiene students (p=.001). 6. Just a small number of the dental hygienists and students knew about the hepatitis B-related past experiences of their families, and the gap between the two was insignificant.
Purpose: Automated system that immunoassay examination are used widely. However, sample to sample carryover can cause that the next patient sample is false positive. Materials and Methods: We test HBs Ag, HBs Ab, HBc Ab(IgG) with Automated pipetting system (Tecan). It was placed with very high concentrations followed by saline solution. During this experiment, The fixed tip of Automated system wash With 0.25 moL/L NaOH. The Measurement results of saline solution confirm the occurrence of carryover. Results: Results of saline solution with 0.25 moL/L NaOH cleaning process was measured 100% negative, And results of patient serum with 0.25 moL/L NaOH cleaning process is similar reported results. Conclusion: As Results, 0.25 moL/L NaOH cleaning process was avoid carryover in experiment, And we know results of the hepatitis test did not affected by this solution we recommend 0.25 moL/L NaOH cleaning process as the Prevention of carryover in the automated system with fixed Tips.
To estimate the prevalence of hepatitis B virus (HBV) and hepatitis C virus (HCV) infection and to determine associated risk factors, a population-based seroepidemiologic study was carried out. In 1993, a health examination survey of the population was carried out in rural area known to have a high incidence of liver cancer. The study population were those who volunteered to participate in a health survey over 10 years of age. Examinees were interviewed by specially trained staffs. Sera from 1,033 study subjects were tested for hepatitis B surface antigen (HBsAg) by .everse passive hemagglutinin (RPHA) estimation and for hepatitis C virus antibody (anti-HCV) by 2nd generation passive hemagglutinin (PHA) estimation. The age and sex standardized prevalence of HBsAg was 6.3% which was similar to national average, but that of anti-HCV was 5.1% which was 4 to 5 times higher than that of blood donors or other health examinees in Korea. In a multivariate analysis, transfusion history, surgical operative history, and acupuncture history were not associated with HBsAg positivity. In contrast, acupuncture history (adjusted odds ratio[OR]=2.2 : 95% Confidence interval[CI] 1.0-4.7) and surgical operative history(adjusted OR=2.0 : 95% CI 1.0-4.1) were associated with anti-HCV positivity. The present study suggest that there is an highly endemic area of HCV infection in Korea and probably this endemicity is associated with a parenteral source of HCV infection other than blood transfusion.
To evaluate the significance of serum $beta_2-microglobulin$ in patients with various liver diseases, serum $\beta_2m$ levels were measured in 44 cases of normal controls, 32 cases of asymptomatic HBsAg carriers and 134 patients with various liver diseases, by radioimmunoassay using Phadebas $Beta_2-micro$ test kits. The following results were obtained: 1) The mean level of serum $\beta_2m$ was $1.39{\pm}0.25mg/l(Mean{\pm}S.D.)$ in normal controls ($1.39{\pm}0.23mg/l$ in 24 males, $1.38{\pm}0.27mg/l$ in 20 females). 2) The serum levels of $\beta_2m$ in patients with various liver diseases and asymptomatic HBsAg carriers were as follows; $1.40{\pm}0.27mg/l$ in asymptomatic HBsAg carriers, $2.42{\pm}0.37mg/l$ in 45 patients with acute viral hepatitis, $2.10{\pm}0.26mg/l$ in 46 patients with chronic persistent hepatitis, $2.60{\pm}0.34mg/l$ in 23 patients with chronic active hepatitis, and $2.60{\pm}0.49mg/l$ in 20 patients with liver cirrhosis. Serum $\beta_2m$ levels of each disease group were significantly higher than that of normal controls(p<0.001). 3) There was significant correlation between the levels of serum $\beta_2m$ and the degrees of lymphocytic infiltration in patients with chronic active hepatitis(p<0.001). 4) Significant correlations were observed between the levels of serum $beta_2-microglobulin$ and serum alanine aminotransferase(r=0.68, p<0.05) and bilirubin(r=0.63, p<0.05) in 15 patients with acute viral hepatitis. In conclusion, the serum $beta_2-microglobulin$ levels were increased in patients with various liver diseases, and it may serve as a new index of liver disease activity.
The single nucleotide polymorphism (SNP) ss469415590 in the interferon lambda-4 (IFNL4) gene has recently been reported to have an association with treatment response in chronic hepatitis C. However, any importance of the SNP in association with response to pegylated interferon (PEG-IFN) therapy in patients with chronic hepatitis B (CHB) is unclear. We retrospectively analyzed data for Thai patients with CHB treated with PEG-IFN for 48 weeks. Virological response (VR) for HBeAg-positive CHB was defined as HBeAg seroconversion plus HBV DNA level <2,000 IU/mL at 24 weeks post-treatment. VR for HBeAg-negative CHB was defined as an HBV DNA level <2,000 IU/mL at 48 weeks. The SNP was identified by real time PCR using the TaqMan genotyping assay with MGB probes. A total 254 patients (107 HBeAg-positive and 147 HBeAg-negative) were enrolled in the study. The distribution of TT/TT, ${\Delta}G/TT$ and ${\Delta}G/{\Delta}G$ genotypes was 221 (87.0%), 32 (12.6%) and 1 (0.4%), respectively. Patients with non-TT/TT genotypes had significantly higher baseline HBV DNA levels than patients with the TT/TT genotype. In HBeAg-positive CHB, 41.2% of patients with TT/TT genotype versus 50.0% with non-TT/TT genotype achieved VR (P=0.593). In HBeAg-negative CHB, the corresponding figures were 40.3% and 43.5%, respectively (P=0.777). There was no significant correlation between the SNP genotypes and HBsAg clearance in both groups of patients. In summary, ss469415590 genotypes were not associated with response to PEG-IFN in Thai patients with HBeAg-positive and HBeAg-negative CHB.
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