• Journal of Life Science
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• v.23 no.2
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• pp.175-181
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• 2013

In Korea, most hepatitis A virus is the IA genotype, but reports of other genotypes have increased recently. Therefore, the purpose of this study is to conduct a genotypic analysis of acute hepatitis A virus. From April 2010 to April 2011, clinical specimens from 20 patients hospitalized with acute hepatitis A and 36 sera positive for anti-HAV IgM were obtained, and the genotype of the VP1/P2A region was analyzed. RNA sequences of the VP1/P2A junction region were amplified using RT-PCR, and the sequences were compared. From 50 sequences amplified, 4 sequences (8%) belonged to genotype IA. The remaining 46 (92%) belonged to genotype IIIA. The results indicate that the genotype of the hepatitis A virus has changed from IA to IIIA in Korea.

• BMB Reports
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• v.38 no.4
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• pp.399-406
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• 2005

A sensitive method for detection of Hepatitis A virus (HAV) by utilizing gold-DNA probe on an array was developed. Amino- modified oligodeoxynucleotides at the 5' position were arrayed on an activated glass surface to function as capture probes. Sandwich hybridization occurred among capture probes, the HAV amplicon, and gold nanoparticle-supported oligonucleotide probes. After a silver enhancement step, signals were detected by a standard flatbed scanner or just by naked eyes. As little as 100 fM of HAV amplicon could be detected on the array. Therefore, the array technology is an alternative to be applied in detection of HAV due to its low-cost and high-sensitivity.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.4 no.2
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• pp.191-198
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• 2001

Purpose: This study was undertaken to investigate the seroepidemiologic pattern of Helicobacter pylori (H. pylori) and hepatitis A virus (HAV) infections in children. Methods: A total of 315 serum samples were obtained from healthy children, living in Gwangju and Chonnam area. All serum samples were assayed for H. pylori IgG level using enzyme immunoassay techniques. HAV IgG level in serum were tested by a competitive radio-immunoassay in 215 subjects. The age-specific seroprevalence of H. pylori and HAV was separately analysed. The concordance of seropositivity and seronegativity between H. pylori and HAV infection was examined by the kappa statistic analysis. Results: Seropositivity was found in 17.5% (55/315) and 30.2% (65/215) of the subjects for H. pylori and HAV, respectively. Cross-tabulation of these data showed that 21 subjects (9.8%) were seropositive and 135 (62.8%) were seronegative for both H. pylori and HAV, 15 (7.0%) were seropositive for only H. pylori and 44 (20.5%) for only HAV. The seroprevalence of H. pylori and HAV increased significantly with age. There was a slight agreement between H. pylori and HAV seropositivity (${\kappa}$=0.26). Conclusion: This study shows a slight similarity in the concordance of seropositivity and seronegativity between H. pylori and HAV infection and provides evidence that H. pylori and HAV may share a common mode of transmission.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.14 no.1
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• pp.59-66
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• 2011

Purpose: We previously reported that concurrent reactivation of latent Epstein-Barr virus (EBV) in children with hepatitis A virus (HAV) infection is common and EBV reactivation with HAV infection adversely affects the clinical features of hepatitis. However, the incidence of concurrent reactivation was not accurate because the detection of EBV reactivation was based on serologic methods. Therefore, we studied the effects of polymerase chain reaction (PCR)-proven EBV reactivation, thus a more precise concurrence, on acute HAV infection in children. Methods: PCR were conducted in 34 patients, who had enrolled previous study and diagnosed with acute HAV infection between January 2008 and June 2010. Their medical records were reviewed. Results: Among 34 patients with acute HAV infection, 12 patients (35.3%) had EBV reactivation which was proven using serologic and molecular biologic techniques. There were significant differences in the peak levels of AST and ALT between the reactivated and non-reactivated groups (p=0.001 and p<0.001, respectively). The duration of full recovery from hepatitis was more prolonged in the reactivated group (p<0.001). Clinical parameters, such as serum protein (p<0.001) and albumin concentrations (p<0.001), atypical lymphocyte count (p=0.001), prothrombin time-international normalized ratio (PT-INR, p<0.001), and splenomegaly (p<0.001), showed significant differences. The clinical features in the reactivated sub-group >10 years of age revealed more liver dysfunction compared to the non-reactivated sub-group. A comparison with a previous study was performed. Conclusion: PCR-proven reactivation of latent EBV in children with HAV infection is common and EBV reactivation with HAV infection adversely affects the clinical features of hepatitis, especially in older children.

• Journal of Microbiology and Biotechnology
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• v.18 no.7
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• pp.1317-1325
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• 2008

Viral safety is an important prerequisite for clinical preparations of plasma-derived pharmaceuticals. One potential way to increase the safety of therapeutic biological products is the use of a virus-retentive filter. In order to increase the viral safety of human antihemophilic factor IX, particularly in regard to non-enveloped viruses, a virus removal process using a polyvinylidene fluoride membrane filter (Viresolve NFP) has been optimized. The most critical factor affecting the filtration efficiency was operating pH and the optimum pH was 6 or 7. Flow rate increased with increasing operating pressure and temperature. Recovery yield in the optimized production-scale process was 96%. No substantial changes were observed in the physical and biochemical characteristics of the filtered factor IX in comparison with those before filtration. A 47-mm disk membrane filter was used to simulate the process performance of the production-scale cartridges and to test if it could remove several experimental model viruses for human pathogenic viruses, including human hepatitis A virus (HAV), porcine parvovirus (PPV), murine encephalomyocarditis virus (EMCV), human immunodeficiency virus type 1 (HIV), bovine viral diarrhea virus (BVDV), and bovine herpes virus (BHV). Non-enveloped viruses (HAV, PPV, and EMCV) as well as enveloped viruses (HIV, BVDV, and BHV) were completely removed during filtration. The log reduction factors achieved were $\geq$6.12 for HAV, $\geq$4.28 for PPV, $\geq$5.33 for EMCV, $\geq$5.51 for HIV, $\geq$5.17 for BVDV, and $\geq$5.75 for BHV. These results indicate that the virus filtration process successfully improved the viral safety of factor IX.

• Microbiology and Biotechnology Letters
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• v.37 no.4
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• pp.377-382
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• 2009

Viral safety is an important prerequisite for clinical preparations of all biopharmaceuticals derived from plasma, cell lines, or tissues of human or animal origin. To ensure the safety, implementation of multiple viral clearance (inactivation and/or removal) steps has been highly recommended for manufacturing of biopharmaceuticals. Of the possible viral clearance strategies, Ultraviolet-C (UVC) irradiation has been known as an effective viral inactivating method. However it has been dismissed by biopharmaceutical industry as a result of the potential for protein damage and the difficulty in delivering uniform doses. Recently a continuous flow UVC reactor (UVivatec) was developed to provide highly efficient mixing and maximize virus exposure to the UV light. In order to investigate the effectiveness of UVivatec to inactivate viruses without causing significant protein damage, the feasibility of the UVC irradiation process was studied with a commercial therapeutic protein. Recovery yield in the optimized condition of $3,000\;J/m^2$ irradiation was more than 98%. The efficacy and robustness of the UVC reactor was evaluated with regard to the inactivation of human immunodeficiency virus (HIV), hepatitis A virus (HAV), bovine herpes virus (BHV), bovine viral diarrhea virus (BVDV), porcine parvovirus (PPV), bovine parvovirus (BPV), minute virus of mice (MVM), reovirus type 3 (REO), and bovine parainfluenza virus type 3 (BPIV). Non enveloped viruses (HAV, PPV, BPV, MVM, and REO) were completely inactivated to undetectable levels by $3,000\;J/m^2$ irradiation. Enveloped viruses such as HIV, BVDV, and BPIV were completely inactivated to undetectable levels. However BHV was incompletely inactivated with slight residual infectivity remaining even after $3,000\;J/m^2$ irradiation. The log reduction factors achieved by UVC irradiation were ${\geq}3.89$ for HIV, ${\geq}5.27$ for HAV, 5.29 for BHV, ${\geq}5.96$ for BVDV, ${\geq}4.37$ for PPV, ${\geq}3.55$ for BPV, ${\geq}3.51$ for MVM, ${\geq}4.20$ for REO, and ${\geq}4.15$ for BPIV. These results indicate that UVC irradiation using UVivatec was very effective and robust in inactivating all the viruses tested.

• Microbiology and Biotechnology Letters
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• v.38 no.2
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• pp.168-176
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• 2010

Acellular dermal matrix (ADM), produced by decellularization from human cadaveric skin, has been used for various biomedical applications. A manufacturing process for ADM ($SureDerm^{TM}$) using tri-n-butyl phospahate (TnBP) and deoxycholic acids as the decellularization solution has been developed. The manufacturing process for $SureDerm^{TM}$ has 70% ethanol treatment and ethylene oxide gas sterilization for inactivating infectious microorganisms. The purpose of this study was to examine the efficacy of the 70% ethanol treatment, decellularization process using 0.1% TnBP and 2% deoxycholic acids, and EO gas sterilization process in the inactivation of viruses. A variety of experimental model viruses for human pathogens, including the human immunodeficiency virus type 1 (HIV-1), bovine herpes virus (BHV), bovine viral diarrhoea virus (BVDV), hepatitis A virus (HAV), and porcine parvovirus (PPV) were all selected for this study. Enveloped viruses such as HIV-1, BHV, and BVDV were effectively inactivated to undetectable levels by 70% ethanol treatment. However HAV and PPV showed high resistance to 70% ethanol treatment with the log reduction factors of 1.85 and 1.15, respectively. HIV-1, BHV, and BVDV were effectively inactivated to undetectable levels by decellularization process. All the viruses tested were completely inactivated to undetectable levels by EO gas treatment. The cumulative log reduction factors of HIV-1, BHV, BVDV, HAV, and PPV were $\geq12.71$, $\geq18.08$, $\geq14.92$, $\geq6.57$, and $\geq7.18$, respectively. These results indicate that the production process for $SureDerm^{TM}$ has a sufficient virus-reducing capacity to achieve a high margin of the virus safety.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.15 no.2
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• pp.91-99
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• 2012

Purpose: The aims of this study was to compare and evaluate the clinical characteristics, laboratory data, and prognosis for infants under age 1 year with CMV hepatitis and those with viral hepatitis of unknown etiology. Methods: A retrospective study was conducted of infants under age 1 year who were admitted with acute hepatitis. The exclusion criteria consisted of: autoimmune, genetic, metabolic, toxic, HAV, HBV, HCV, toxoplasma, rubella, herpes simplex, and Epstein-Barr virus. The 30 patients included were divided into two groups based on markers for CMV (IgM anti-CMV, CMV PCR in urine, CMV culture in urine). Results: The median age of patients (n=15) was 2.8 months. No other organ involvement was detected in any patient. Peak serum total bilirubin levels (n=4) ranged from 2.6 to 6.7 mg/dL. Peak serum ALT levels ranged from 51 to 1,581 IU/L. The duration of ALT elevation ranged from 1.5 weeks to 26 weeks (median 9 weeks). All had recovered in full without ganciclovir; there were no cases of hearing loss. The median age of controls (n=15) was 2.5 months. Peak serum total bilirubin levels (n=4) ranged from 1.6 to 9.1 mg/dL. Peak serum ALT levels ranged from 26 to 1,794 IU/L. No significant differences were observed between both groups regarding the peak serum ALT levels, peak serum total bilirubin levels, duration of hyperbilirubinemia and ALT elevation. Conclusion: Although it was not possible to differentiate congenital infection with perinatal infection in this study, the prognosis of patients with CMV hepatitis without other organ involvement was good without ganciclovir treatment.

• Journal of Life Science
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• v.12 no.1
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• pp.49-54
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• 2002

Development of a rapid method possessing the requisite sensitivity and specificity for virus monitoring is necessary for protection of the shellfish-consuming public. Oysters tissue usually contains virus particles in relatively small concentrations along with various other substances that can interfere with detection steps. Therefore, the critical point concerning the detection of viruses in shellfish tissues resides in the processing of samples. The current study demonstrated the possibility of purifying small amounts of virus particles at the interface of a 10/50% sucrose gradient after a single round of sucrose gradient ultracentrifugation. We could detect HAV and poliovirus simultaneously from oyster tissues by using two different sets of primer. Furthermore, the method showed a high level of virus recovery rate (>95%) as determined by plaque assays of the final samples. Taken the advantages of the simple and sensitive methods, it was possible to detect 2 pfu of HAV in 5 g of oyster digestive tissues within 24h.

• Pediatric Infection and Vaccine
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• v.4 no.2
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• pp.232-239
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• 1997

Purpose: The incidence of hepatitis A virus(HAV) infection has markedly decreased in the last 20 years due to industrialization and improvements in the standard of living and hygiene in Korea. The reduction in seroprevalence rates indicates infection potential for young adult population, and a need for vaccinations in high-risk adults and children groups has been suggested. In this study we evaluated the seroprevalence rates and natural infection rates of hepatitis A in children and adolescent to obtain the basic data for vaccination of hepatitis A. Methods: A total of 334 children and adolescent subjects below 20 years old in Kyonggi-do province were examined for HAV antibody and seroprevalence rates in each age group was investigated. In 584 elementary school students residing in Kyonggi-do province, serum samples collected in 1993 and 1996 from the same subjects for investigation of natural seroconversion rates. Method of testing antibody was enzyme immunoassay. Results: 1) The seropositive rate of HAV in 334 subjects aged below 20 years old was 5.4%. According to age, the seropositive rates were 27.3% in infant group, 0.0% in 1~4 year-old group, 0.0% in 5~9 year-old group, 2.9% in 10~14 year-old group and 15.0% in 15~19 year-old group. 2) In the study of 584 elementary school children, only one subject showed seropositive in 1993, and in 1996 three different subjects showed seropositive results(0.5%); the natural seroconversion rate during 3 years was 0.5%. 3) The seroprevalence rates of below 20 year-old subjects reported in previous studies were 63.8% in 1979 and 47.3% in 1989 while the present study showed the rate at 5.4%. Conclusion: Since natural antibody formation is rarely occurring, there is a high risk for apparent hepatitis A infection in adults. Therefore vaccination in high risk groups is essential at present, and in order to reduce the chance for hepatitis A infection in adults, vaccination in children may be needed.