As part of the immunoserology program of the Korean Association of External Quality Assessment Service, we organized two trials on the external quality assessment of hepatitis viral markers in 2016 and 2017. The hepatitis viral antigens and antibodies program consisted of 10 test items. We delivered two and three types of pooled sera specimens to 965 and 965 institutions for the first and second trials of external proficiency testing in 2016, respectively. The number of participating laboratories was 915 (94.8%) and 913 (95.0%) in the first and second trials in 2016, respectively. We also delivered three kinds of pooled sera specimens to 936 and 1,015 institutions for the first and second trials of external proficiency testing in 2017, respectively. The number of participating laboratories was 920 (98.3%) and 996 (98.1%) in the first and second trials in 2017, respectively. The most commonly tested items were hepatitis B surface antigen, followed by the antibodies to hepatitis B surface antigen, anti-hepatitis C virus, hepatitis B envelope antigen, antibodies to hepatitis B envelope antigen, anti-hepatitis A virus and antibodies to hepatitis B core antigen. The most frequently used methods for detecting viral markers were the chemiluminescence immunoassay and the electrochemiluminescence immunoassay, but they yielded a few-false positive results due to the matrix effect. The immunochromatographic assay yielded false-negative results for anti-hepatitis A virus due to low sensitivity. Continuous improvement in the quality of viral hepatitis testing through participation in the survey seems necessary.
Antibodies ; Antigens, Viral ; Biomarkers* ; Hepatitis A ; Hepatitis B ; Hepatitis B Core Antigens ; Hepatitis B Surface Antigens ; Hepatitis B virus ; Hepatitis C ; Hepatitis* ; Immunoassay ; Immunochromatography ; Korea* ; Laboratory Proficiency Testing ; Luminescence

<b>OBJECTIVEb>To establish a national reference panel for HIV RNA diagnostic reagents.

<b>METHODSb>Sera from patients with HIV infection and healthy blood donors were collected and tested for HIV and HCV antibodies and HBsAg by using ELISA. The HIV antibody positive samples with ELISA were confirmed with HIV Blot 2.2 (Genelabs). The quantitative samples for HIV RNA were calibrated with the WHO HIV RNA standard. The stability of the panel was evaluated with acceleration method.

<b>RESULTSb>After screening and calibration, 8 negative samples, 8 positive samples, 3 quantitative samples, 6 sensitivity samples and 5 samples for linear analysis were composed of the national reference panel for HIV RNA. The convinced international units (IU) for the quantitative samples were obtained by seven independent calibration and the logarithm of international units for the quantitative samples (b1-b3) were less than x +/- s. The results showed that this panel may stabilize for 4 days at 4 degrees C.

<b>CONCLUSIONb>A national reference panel for HIV RNA reagents has been established. It may provide the basis for evaluating HIV RNA diagnostic reagents.

Blood Donors ; Calibration ; Drug Stability ; HIV Antibodies ; blood ; HIV Infections ; blood ; virology ; HIV-1 ; genetics ; isolation & purification ; Hepatitis B Surface Antigens ; blood ; Hepatitis C Antibodies ; blood ; Humans ; Indicators and Reagents ; standards ; RNA, Viral ; standards ; Reference Standards ; Reproducibility of Results ; Sensitivity and Specificity

<b>OBJECTIVEb>To analyse the proportion of hepatitis associated aplastic anemia (HAAA) in severe aplastic anemia (SAA) and its clinical features of HAAA.

<b>METHODSb>All newly diagnosed SAA cases in our department in the recent 5 years were analyzed. A case-control study was undertaken to investigate the differences of clinical and laboratory features between HAAA and non-hepatitis associated SAA (non-HASAA) patients.

<b>RESULTSb>The proportion of HAAA in SAA was 3.3%. There was no significant difference in PB cell counts, bone marrow hematopoiesis status and the amount of blood transfusion between HAAA and non-HASAA patients. Sera from 13 patients with HAAA were tested for antibodies to hepatitis viruses A, B, and C and hepatitis B surface antigen. Twelve (92.3%) of them had negative serologic results for the tests and only one (7.7%) had a positive result for HBsAg and HBeAg. Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were decreased prior to the diagnosis in twelve (92.3%) of the 13 HAAA patients. The percentage of CD4(+) cells in HAAA patients was significantly lower than that in non-HASAA patients (P < 0.05). HAAA patients had higher percentages of CD8(+) cells (P < 0.05) and lower ratios of CD4(+)/CD8(+) (P < 0.05). The early infection rate of the HAAA patients was significantly higher than that of non-HASAA patients (84.6% vs 42.3%, P < 0.05), with different mortalities (61.5% vs 15.4%, P < 0.05). The 2-year survival rate of HAAA patients was significantly lower than that of non-HASAA patients (16.6% vs 83.2%, P < 0.01).

<b>CONCLUSIONb>The proportion of HAAA in SAA was 3.3%. Most of HAAA were associated with non-A, non-B and non-C hepatitis virus. Compared with that of non-HASAA, the abnormality of T cell immunity of HAAA was more severe, with a higher frequency of early infection and a higher mortality rate.

Adolescent ; Adult ; Anemia, Aplastic ; blood ; complications ; pathology ; Case-Control Studies ; Female ; Follow-Up Studies ; Hepacivirus ; immunology ; Hepatitis A Antibodies ; blood ; Hepatitis A virus ; immunology ; Hepatitis B Antibodies ; blood ; Hepatitis B virus ; immunology ; Hepatitis C Antibodies ; blood ; Hepatitis, Viral, Human ; blood ; complications ; virology ; Humans ; Male

Hepatitis viruses (hepatitis B virus (HBV) and hepatitis C virus) have been associated with development of inflammatory arthritis. Approximately 400 million people worldwide have chronic HBV infection. HBV infection is the one of the most common causes of liver disease, and the prevalence of HBV infection in Korea is almost 6%. Arthritis in patients with HBV can be encountered in two settings: as a rheumatoid arthritis (RA)-like, acute, self-limited polyarthritis during the pre-symptomatic phase of acute hepatitis B, or, more rarely, as arthritis occurring in the context of HBV-associated polyarteritis nodosa (PAN). In both cases, the pathogenesis of arthritis is attributed to the deposition of immune complexes containing viral antigens (HBsAg or HBeAg) and their respective antibodies (anti-HBs and anti-HBe) in synovial tissues. Here we report on a case of polyarthritis associated with reactivation of chronic hepatitis B virus infection with a review of the literature.
Antibodies ; Antigen-Antibody Complex ; Antigens, Viral ; Arthritis* ; Arthritis, Rheumatoid ; Hepatitis B ; Hepatitis B, Chronic* ; Hepatitis C ; Hepatitis Viruses ; Hepatitis, Chronic* ; Herpesvirus 1, Cercopithecine ; Humans ; Korea ; Liver Diseases ; Polyarteritis Nodosa ; Prevalence ; Viruses

<b>OBJECTIVEb>To investigate the seroprevalence of hepatitis viruses in Mianyang of the Sichuan province.

<b>METHODSb>EIISA was used for detecting anti-HAV IgG, HBsAg/HBsAb, anti-HCV IgG and anti-HEV IgG of the serum samples. All sample were collected in Mianyang areas in 2007.

<b>RESULTSb>1352 samples were detected. The positive rates of anti-HAV, HBsAg/HBsAb, anti-HCV,and anti-HEV are 81.07% (1096/1352), 5.40% (73/1352) and 61.32% (829/1352), 0.37% (5/1352) and 49.26% (666/1352), respectively. The positive rate at different age group, for anti-HAV was 38.21% of 10-19 years old, 83% of 20-29 years old, 88% of 30-39 years old, 95.03% of 40-49 years old, 97% of 50-59 years old, 97.77% of 60-69 years old, 97.52% of > or =70 years old. For HBsAg/HBsAb were 5.65% or 50.83%, 10.0% or 68.0%, 5.20% or 78.80%, 5.97% or 78.11%, 6.50% or 62.50%, 1.12% or 51.40%, 4.96% or 30.58% at the same age group, respectively,for anti-HCV, was 0.33% of 10-19 years old, 0.80% of 30-39 years, 0.56% of 60-69 years old, 0.83% of > or =70 years old.For HEV-IgG was 26.58% of 10-19 years old, 42.0% of 20-29 years old, 55.22%-61.0% of 30-> or =70 years old, for anti-HEV IgM, was 10.06% (53/527) in the positive samples of HEV-IgG.

<b>CONCLUSIONb>The inoculation againt HAV and HBV is enhanced in the young population. HBsAg carrier and HCV infection is decreasing. The HEV infection is actually increasing.

Adolescent ; Adult ; Aged ; Antibodies, Anti-Idiotypic ; blood ; Antibodies, Viral ; blood ; Child ; China ; epidemiology ; Female ; Hepacivirus ; immunology ; isolation & purification ; Hepatitis A ; epidemiology ; immunology ; Hepatitis Antibodies ; blood ; classification ; Hepatitis B ; epidemiology ; immunology ; Hepatitis B virus ; immunology ; isolation & purification ; Hepatitis C ; epidemiology ; immunology ; Hepatovirus ; classification ; immunology ; isolation & purification ; Humans ; Immunoglobulin M ; blood ; Male ; Middle Aged ; Seroepidemiologic Studies ; Young Adult

<b>BACKGROUNDb>To study preparation of polyvalent DNA vaccine and the control of multiple gene expression.

<b>METHODSb>A bicistronic vector pcDNA3.0BA was constructed from pcDNA3.0. HCV PC154 gene and HBV preS2S gene were inserted into this vector to form bicistronic expression construct pcDNA3.0BAPC154S2S and monocistronic expression construct pcDNA3.0BAPC154 or pcDNA3.0BAS2S. These plasmids were transiently expressed in COS-7 cells and injected into muscles of BALB/c mice.

<b>RESULTSb>pcDNA3.0BA contains two cistronic units, which can co-express two kinds of genes, with the first immunogen gene and the second gene serving as additional immunogen or as modulator for the immune responses. HBV surface Ag and HCV core Ag were coexpressed in vitro. The antibody responses and lymphoproliferation to antigens were similar between bicistronic and monocistronic expression construct in mice.

<b>CONCLUSIONb>pcDNA3.0BA is a novel vector, which can coexpress two proteins and elicit polyvalent immune responses.

Animals ; COS Cells ; Cercopithecus aethiops ; DNA, Recombinant ; immunology ; Gene Expression ; Hepatitis B ; blood ; immunology ; Hepatitis B Antibodies ; blood ; Hepatitis B Surface Antigens ; genetics ; immunology ; Hepatitis C ; blood ; immunology ; Hepatitis C Antibodies ; blood ; Hepatitis C Antigens ; genetics ; immunology ; Immunization ; methods ; Mice ; Mice, Inbred BALB C ; Plasmids ; genetics ; Vaccines, DNA ; genetics ; immunology ; Viral Hepatitis Vaccines ; genetics ; immunology

<b>OBJECTIVEb>To compare the various combined immunization schemes available for treatment of babies born to mothers with high-load hepatitis B virus (HBV) infection.

<b>METHODSb>A total of 118 mothers with HBV infection status of hepatitis B surface antigen-positive (HBsAg+), hepatitis B e antigen-positive (HBeAg+) and HBV DNA load of more than 1.0 * 61og10 IU/mL were included in the study. All of the participants' babies received the main-passive immunization therapy according to the wishes of their families. For analysis,the infants were grouped according to the various dosages of the vaccine program (group A: hepatitis B immunoglobulin (HBIG) 200 IU and HBVac 20 mug intramuscular;group B:HBIG 200 IU and HBVac 10 mug intramuscular; group C HBIG 100 IU and HBVac 20 mug intramuscular injection) and times, and followed-up to 7 months of age.All results were statistically analyzed using SPSS software.

<b>RESULTSb>All of the infants produced anti-HBs after vaccination.After the HBIG injection schedule was completed in January, the mean concentrations of anti-HBs in groups A, B, and C were 263.56 ± 50.98,231.06 ± 74.07, and 99.23 ± 29.82 mIU/mL respectively;the concentrations were significantly different between groups A and C, and between groups B and C (P < 0.001). In July, the titers of anti-HBs in groups A, B, and C were 788.10 ± 281.96,428.39 ± 347.48, and 708.44 ± 315.69 mIU/mL respectively; the concentrations were significantly different between groups A and B, and between groups B and C (P < 0.05).

<b>CONCLUSIONb>AdminisWation of the hepatitis B vaccine combined with HBIG at birth can achieve immune protection for babies born to highly viremic mothers. In January, the HBIG dosage of 200 IU was more reliable than 100 IU. The hepatitis B 20 tg dose vaccine was safe and effective.

Hepatitis B ; Hepatitis B Antibodies ; Hepatitis B Vaccines ; Hepatitis B e Antigens ; Hepatitis B virus ; Humans ; Immunization ; Immunoglobulins ; Infant ; Mothers ; Serologic Tests ; Vaccines, Combined ; Viral Load

Human antibodies generated by phage antibody technology have been widely used in the immunotherapy of various diseases. Among the characteristics of these therapeutic antibodies, affinity is one of the most important determinants of their biological efficacy. The binding of an antibody and its corresponding antigen could be disrupted by thiocyanate solution of different concentrations, depend upon the affinity of the antibody. This mechanism has been adopted to determine the relative affinity of monoclonal or polyclonal antibodies in routine immunological practice. Correlation between the elution method and other techniques that measure the affinity such as equilibrium dialysis and biospecific interaction analysis (BIA) has been established. Here we describe the applications of the thiocyanate elution method in the determination of the relative affinity index (RAI) of phage antibodies (Phabs). Five clone antibodies, including 3 clones of anti-keratin antibodies (AK1, AK2 and AK3) and 2 clones of anti-HBsAg antibodies (HB1 and HB2) were selected to express Phabs and Fabs, and the RAI were determined by ELISA after thiocyanate elution. A HRP-conjugated anti-M13 was used as secondary antibody for Phabs and HRP-goat-anti-human Fab was used for Fabs. The affinity ranks of the Phabs were compared with that of the Fab fragments. The results showed that all the Phabs tested were tolerant to thiocyanate treatment. The relative affinity rank of 5 Phabs coincided well with that of their corresponding Fabs. We conclude that the thiocyanate elution can be used as an easy and rapid method to measure and compare the relative affinity of Phabs.
Antibodies ; immunology ; Antibodies, Monoclonal ; immunology ; Antibodies, Viral ; immunology ; Antibody Affinity ; Cloning, Molecular ; Enzyme-Linked Immunosorbent Assay ; methods ; Hepatitis B Antibodies ; immunology ; Hepatitis B Surface Antigens ; immunology ; Keratins ; immunology ; Peptide Library ; Proteomics ; methods ; Thiocyanates ; chemistry ; Transfection

BACKGROUND/AIMS: We studied the prevalence of chronic hepatitis B, C and abnormality on liver function among the population in northwest area of Chungnam. METHODS: We have reviewed 40,112 adults who had received medical examination at health promotion center in Dankook university hospital. We studied them retrospectively about HBsAg, HBsAb, anti-HCV, and liver function test (LFT). RESULTS: Among the study subjects, 22,936 men and 17,176 women were involved. The overall seroprevalence of HBsAg was 4.2%. The prevalence in men (4.5%) was higher than that of women (3.7%) (p<0.001). The seroprevalence of HBsAg in their age was 5.1% in the 5th decade, 4.2% in the 2nd decade, 4.1% in the 4th decade, and 4.1% in the 6th decade. The overall seroprevalence of HBsAb was 65.1%. The overall seroprevalence of anti-HCV was 0.7%. After we reexamined them with HCV RNA or RIBA (Recombinant Immunoblot Assay), the prevalence of chronic hepatitis C was 0.09%. The LFT abnormality in total subjects was 11.4%. The LFT abnormality of chronic hepatitis B and C subjects was 21.72% and 63.2%. CONCLUSIONS: The prevalence of chronic hepatitis B and C was lower than that of previous studies. The prevalence of chronic hepatitis B in the 2nd decade was still high.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Female ; Hepatitis B Antibodies/blood ; Hepatitis B Surface Antigens/blood ; Hepatitis B, Chronic/*epidemiology/immunology/physiopathology ; Hepatitis C Antibodies/blood ; Hepatitis C, Chronic/*epidemiology/immunology/physiopathology ; Humans ; Korea ; Liver Function Tests ; Male ; Middle Aged ; Prevalence ; RNA, Viral/blood ; Retrospective Studies

Occult HBV infection is characterized by the absence of serum HBsAg with persistence of low level of intrahepatic HBV DNA. Several suggested mechanisms for the origin of occult HBV infection include strong suppression of viral replication and gene expression, mutation in the regulatory regions of HBV genome, formation of immunoglobulin-bound HBsAg, viral interference, and blockage of HBsAg secretion from infected hepatocytes. Standardized assays are not yet available, and sensitive HBV DNA amplification assay is necessary for the diagnosis of cryptic infection. Detection rate of HBV DNA is highest in IgG anti-HBc positive population. However, neither anti-HBc nor anti-HBs can be detected in a significant proportion of infected persons. Occult HBV infection occurs in a number of clinical settings and is highly prevalent in HCV-infected patients as well as in patients with cryptogenic chronic liver disease including hepatocellular carcinoma.
DNA, Viral/analysis ; Hepatitis B/*diagnosis/*epidemiology/metabolism ; Hepatitis B Antibodies/blood ; Hepatitis B Core Antigens/immunology ; Hepatitis B Surface Antigens/blood ; Humans