몇가지 간질환에 있어서 DNA등 HBV 표지자의 의의

김재식,이원길,서장수,송경은,이중원,박원림,양정선,김영태,김기연,장윤정,이난영,양선문,양수환,홍현숙,김신학,문정숙,오정도 慶北大學校 醫科大學 1998 慶北醫大雜誌 Vol.39 No.2

HBV : PE-038 ; Association of MicroRNA-323b polymorphisms with clearance of hepatitis B virus infection

( Su Jong Yu ),( Jin Wook Kim ),( Jeong Hoon Lee ),( Jung Hwan Yoon ),( Hyo Suk Lee ),( Chung Yong Kim ),( Yoon Jun Kim ) 대한간학회 2012 춘·추계 학술대회 (KASL) Vol.2012 No.1

Background: Recently, some reports have shown that some mammalian microRNAs (miRNAs) play a role in antiviral defense. However, little is known about the role of miRNA-323b in hepatitis B virus (HBV)-host interaction. We explored whether single nucleotide polymorphism (SNP) of miRNA-323b affect HBV replication or risk of hepatocellular carcinoma (HCC) occurrence in a Korean HBV cohort. Methods: Genotyping was performed in a total of 1,439 subjects composed of 404 spontaneously recovered (SR) subjects as normal controls and 1,035 chronic carriers (CC) of HBV who were further classified into 313 patients with chronic hepatitis (CH), 305 patients with liver cirrhosis (LC), and 417 patients with HCC. To confirm the effect of SNP of miRNA- 323b on HBV replication in vitro, HepAD38 cells were transduced with miRNA-323b wild type or miRNA-323b SNP plasmid vectors, and HBV replication was induced for 5 days. HBV DNA was isolated and quantified using real-time PCR. Results: The polymorphism rs56103835C>T in the primary miRNA region of miRNA-323b revealed significant minor allele frequency (0.273). rs56103835C>T SNP showed significantly affect clearance of HBV in SR group compared to CC group (OR=1.29, p=0.009 in a codominant model; OR=1.29, p=0.03 in a dominant model; and OR=1.78, p=0.03 in a recessive model). In addition, there was a trend towards enhanced clearance of HBsAg in patients with rs56103835C>T SNP (OR=1.24, p=0.05 in a codominant model). In vitro, the total intracellular HBV DNA content was significantly reduced by miRNA-323b SNP plasmid vector transduction (p = 0.008). Conclusion: The CC homozygote of miRNA-323b was significantly associated with clearance of HBV compared to the CT or TT genotypes among SR subjects infected with HBV. Our findings provide a novel perspective on the role SNP of miRNAs in host-virus interactions in HBV infection.

Knowledge of Hepatitis B Virus Infection among Health Science Students of Pokhara Univ., Gandaki Province, Nepal

( Bishnu Raj Tiwari ),( Suresh Jaiswal ),( Sanju Bastola ),( Bimala Sharma ) 대한간학회 2020 춘·추계 학술대회 (KASL) Vol.2020 No.1

Aims: Hepatitis B virus infection (HBV) is public health problem in Nepal. The prevalence of HBV was found to be below 1% (0.9%), however some subgroup of population showed high prevalence by various studies. The study aimed to determine the knowledge of Hepatitis B virus infection among health science students of Pokhara University, Nepal. Methods: A cross-sectional survey was done among 200 students of health and allied science of Pokhara University using a structured self-administered questionnaire method. The questionnaire was prepared based on the previous studies. Students from public health, laboratory science, pharmacy and nursing disciplines were enrolled in the study. The information was collected from January to February, 2020. Mean knowledge score of HBV was measured based on 28 related questions as it was done in previous study, in which each correct answer was coded 1 and incorrect answer as 0, resulting the maximum score of 28 for all right answers. Descriptive statistics, and chisquare test, independent sample t test and one -way ANOVA test were applied. Results: Of the total, 74.5% of the respondents were females; and mean age of the respondents was 20.6 (SD ±1.48) years. Mean knowledge score of HBV was 19.48 (SD±2.72) out of 28. The mean knowledge score was 19.90 (SD± 2.67) among females and 18.33 (SD±2.53) among males; there was significant difference in the mean knowledge score between males and females (p value <0.05). Similarly, there was also significant difference in the mean knowledge score among the students of different disciplines of health science (p value <0.05); the highest mean was observed among nursing students which was 21.22 (SD+2.64). Regarding the individual questions, 98% respondents agreed that hepatitis B infection is caused by a virus. Among all, 16.5% respondents agreed that hepatitis B infection can be spread by mosquitoes; the statement was significantly different by sex (P<0.05). Of the total, 41.9% agreed that hepatitis B can be spread through close personal contact such as kissing or talking, 29.5% agreed that sharing dishes with HBV positive patients can cause the spread of virus; 97% reported that hepatitis B is spread through blood-to-blood contact; and 86.5% respondents mentioned that sexual transmission is a common way hepatitis B is spread. Of total, 91% students agreed that there is a vaccine for HBV. Among all, 84% students agreed that HBV is associated with an increased risk of liver cancer and 87.5% mentioned that HBV can lead to liver cirrhosis. Of the total, 58.5% agreed that having a medical and/or dental procedure increases a person’s chances of contracting HBV; 69.5% students believed that symptoms appear soon after the entrance of HBV into the body; 55.8% agreed that symptoms always appear after the entrance of HBV to the body; 38.5% students agreed that people with HBV should be restricted from working in the food industry; and 64.5% students reported that there is a pharmaceutical treatment available for hepatitis B; and the agreement with all the above statements had statistically significant association with sex (p value <0.05). Conclusions: The mean knowledge score of HBV infection was found satisfactory among health science students of Pokhara University, Gandaki province, Nepal. Significant differences were observed in the mean knowledge score of HBV by sex and different disciplines of health science. The curriculum of all health science disciplines should be comprehensive to cover all aspect of HBV infection

Comparative Study of Cirrhosis Stage in Patients with HBV Infection and HBV/HDV Co-Infection in Mongolia

( Tsevelmaa Munkhchuluun ),( Dashchirev Munkh-orshikh ),( Baasankhuu Enkhtuvshin ),( Oidov Baatarkhuu ) 대한간학회 2020 춘·추계 학술대회 (KASL) Vol.2020 No.1

Aims: Comparative study of cirrhosis stage in patients with HBV infection and HBV/HDV co-infection Methods: Our study continued from January 2015 to March 2017 and we measured liver fibrosis stage in patients with HBV infection and HBV/HDV co-infection using a Fibroscan (Fibroscan 502 Touch, Echosens, and Paris, France) who are controlled in Happy Veritas Clinic and Diagnostic Center. When we measured liver fibrosis stage in 5504 patients with HBV infection, 20% or 1115 of the patients is determined HDV co-infection. In our study in random sampling cases are selected 354 patients with HBV mono infection and 177 of all patients have HBV/HDV co-infection. We selected parameters from patient’s medical histories in our study, such as serologic markers of HBV quantification of HBV and HDV in serum samples, blood test, liver function tests, and liver fibrosis stage. Results: 354 patients 47.7% (169) was men. Range with an average age of 44±17 (range 18-75 years old) were included the study. According to the comparative study in laboratory tests, ALT level was HBV - 44 (36; 51.5) and HBV/HDV co-infection 61 (39.8; 97.5), AST level was HBV - 39.1(30; 83) and HBV/HDV co-infection - 50 (33.1; 77.8), Platelet count was HBV- 193±66 and HBV/HDV - 181±62.8. When we compared liver fibrosis stage were HBV- F0 67(37.9%), HBV/HDV-F0 57(32.2%), HBV-F1 22(12.4%), HBV/HDV-F1 17(9.6%), HBV-F2 39(22%), HBV/HDV- F2 39(22%), HBV-F3 29(16.4%), HBV/HDV-F3 41(23.2%), HBV- F4 20(11.3%) and HBV/HDV - F4 23(13%) . In table 1 shows the difference of liver fibrosis by age group. Conclusions: 65.5% of all patients with HBV/HDV co-infection are from 30 to 50 years old. Liver fibrosis of patients with HBV/HDV co-infection is a higher 11.88kPa than patients with HBV mono-infection. Our study shows that, the hepatitis is more severe in patients with HBV/HDV co-infection and the platelet count is less than HBV infection only.

HBV : PE-031 ; Occult HBV infection in Korean blood donors

( Ja Kyung Kim ),( Hye Young Chang ),( Myung Hee Kim ),( Hyun Ok Kim ),( Mi Yeon Choi ),( Soo Youn Lee ),( Hyun Chung Chung ),( Yoo Sung Hwang ),( Kwan Sik Lee ) 대한간학회 2012 춘·추계 학술대회 (KASL) Vol.2012 No.1

Background: Blood transfusion can transmit HBV-DNA and cause hepatitis B in blood recipient. Currently, serologic assay for the detection of HBsAg has been used in routine screening test of blood donors. Korea is an endemic area of HBV infection, prevalence of occult hepatitis B can be high. However, donors, who are suspected to have occult HBV infection, have not been evaluated further. We aimed to reveal clinical characteristics and to analyze HBV-DNA in liver tissue in these donors. Methods: Patients who donated blood in Hanmaeum Blood Center between May 16, 2011 and Dec 31, 2011 were enrolled in this study. Blood from these donors was screened HBsAg and HBV-DNA. HBV-DNA was detected by TMA (translationmediated amplification) method with PROCLEIX® TIGRIS® system (Novartis, USA). If a donor, who showed negative HBsAg but positive HBV-DNA, agreed to be evaluated, the donor underwent other serologic tests and liver biopsy. HBV-DNA in liver tissue was detected by direct sequencing. Results: Total 103,017 donors were enrolled. One hundred nineteen donors (0.11%) showed positive HBsAg. Among 102,898 donors with negative HBsAg (99.9%), HBV-DNA was positive in 16 donors (0.016%). Among negative HBsAg and positive HBV-DNA donors, 3 male donors agreed to evaluate further. All three donors had positive anti-HBs and negative HBeAg/anti-HBe, however, only one donor had positive anti-HBc. We confirmed HBV-DNA in liver tissue by PCR. Conclusion: Dual Screening tests with HBsAg and HBV-DNA for blood product can detect occult infection in blood donors. Clinical significance of donors with occult HBV infection needs to evaluated further.

항 바이러스 치료중인 B형 간염환자에서 HBeAg 및 HBV DNA 검출에 관한 분석

천준홍,채홍주,박미선,임수연,유선희,이선호,Cheon, Jun Hong,Chae, Hong Ju,Park, Mi Sun,Lim, Soo Yeon,Yoo, Seon Hee,Lee, Sun Ho 대한핵의학기술학회 2019 핵의학 기술 Vol.23 No.1

[목적] B형 간염바이러스(hepatitis B virus, HBV)감염은 전세계적으로 중요한 공중 보건 문제이며 만성간염, 간 경변, 간암의 주요 원인으로 알려져 있으며, 이러한 질환의 진단 및 치료에 B형 간염바이러스의 혈청학적 검사는 필수적이다. 항 바이러스 치료중인 B형 간염 환자를 대상으로 면역방사계수 측정법(IRMA; Immunoradiometric assay)과 화학발광 미세입자 면역분석법(CMIA; Chemiluminescent Micropartical Immunoassay)을 이용하여 HBe-Ag 검사를 시행하였고, 실시간 중합효소 연쇄반응(RT-PCR; Realtime-Polymerase Chain Reaction)법을 이용하여 혈청 내 HBV DNA 검출율 을 비교 분석 하였다. [대상 및 방법] 항 바이러스 치료가 시행중인 B형 간염 환자 270명을 대상으로 HBeAg 혈청 검사와 HBV DNA 정량 검사를 실시하였다. HBeAg 혈청 검사는 검출 원리가 다른 두 가지 혈청학적 검사법(IRMA, CMIA)을 적용 하였고, 혈청 내 HBV DNA는 Abbott m2000 System을 사용하여 실시간 중합효소 연쇄반응(RT-PCR; Realtime-Polymerase Chain Reaction)법으로 정량 측정 하였다. [결과] HBeAg 검출율은 면역방사계수법(IRMA)의 경우 24.1% (65/205), 화학발광 미세입자 면역 분석법(CMIA)에서는 82.2% (222/48)의 결과를 보였다. 혈청학적 검사방법(IRMA, CMIA)에 따른 HBeAg 검사결과의 일치율은 33% (89/270)이다. 실시간 중합효소 연쇄반응(RT-PCR)을 이용한 혈청 내 HBV DNA의 검출율은 29.3% (79/191)를 보였고, 혈청 내 HBV-DNA 농도는 $16IU/mL{\times}1.0{\times}10^9IU/mL$ 이며 검출한계는 <15IU/mL 이다. 면역방사계수법(IRMA)으로 HBeAg 검출결과가 양성일때 55.4%, 그리고 음성일때 20.9%의 HBV DNA 검출율과 $1.1{\times}10^8IU/mL$, $5.7{\times}10^5IU/mL$의 혈청내 HBV DNA농도를 나타냈다. 이에 반해 화학발광 미세입자 면역 분석법(CMIA)의 경우 HBeAg 검출결과가 양성일때 HBV DNA 검출율은 28.4%, 음성일때 33.3%의 결과를 나타냈으며 혈청 내 HBV DNA농도는 $6.0{\times}10^7IU/mL$, $2.4{\times}10^5IU/mL$ 이었다. 면역방사계수법과 화학발광 미세입자 면역 분석법에서 동일하게 HBeAg 검출 결과가 양성인 경우 HBV DNA 검출율은 62.3%의 결과를 보였으며, 혈청 내 HBV DNA 농도는 $1.1{\times}10^8IU/mL$ 이다. [결론] 혈청학적 검사법에 따른 HeAg 검출율은 많은 차이를 보였다. 이러한 차이는 검사kit에 사용된 Ab의 특성과 epitope, HBV의 genotype등 여러 가지 원인으로 생각된다. 혈청학적 검사 결과로 분류 된 그룹별 HBV DNA의 검출율과 농도를 비교한 결과, Group II(IRMA 양성, CMIA 양성, N=53)에서 높은 검출율과 농도를 확인할 수 있었다. Purpose Hepatitis B virus (hepatitis B virus, HBV) infection is a worldwide major public health problem and it is known as a major cause of chronic hepatitis, liver cirrhosis and liver cancer. And serologic tests of hepatitis B virus is essential for diagnosing and treating these diseases. In addition, with the development of molecular diagnostics, the detection of HBV DNA in serum diagnoses HBV infection and is recognized as an important indicator for the antiviral agent treatment response assessment. We performed HBeAg assay using Immunoradiometric assay (IRMA) and Chemiluminescent Microparticle Immunoassay (CMIA) in hepatitis B patients treated with antiviral agents. The detection rate of HBV DNA in serum was measured and compared by RT-PCR (Real Time - Polymerase Chain Reaction) method Materials and Methods HBeAg serum examination and HBV DNA quantification test were conducted on 270 hepatitis B patients undergoing anti-virus treatment after diagnosis of hepatitis B virus infection. Two serologic tests (IRMA, CMIA) with different detection principles were applied for the HBeAg serum test. Serum HBV DNA was quantitatively measured by real-time polymerase chain reaction (RT-PCR) using the Abbott m2000 System. Results The detection rate of HBeAg was 24.1% (65/270) for IRMA and 82.2% (222/270) for CMIA. Detection rate of serum HBV DNA by real-time RT-PCR is 29.3% (79/270). The measured amount of serum HBV DNA concentration is $4.8{\times}10^7{\pm}1.9{\times}10^8IU/mL$($mean{\pm}SD$). The minimum value is 16IU/mL, the maximum value is $1.0{\times}10^9IU/mL$, and the reference value for quantitative detection limit is 15IU/mL. The detection rates and concentrations of HBV DNA by group according to the results of HBeAg serological (IRMA, CMIA)tests were as follows. 1) Group I (IRMA negative, CMIA positive, N = 169), HBV DNA detection rate of 17.7% (30/169), $6.8{\times}10^5{\pm}1.9{\times}10^6IU/mL$ 2) Group II (IRMA positive, CMIA positive, N = 53), HBV DNA detection rate 62.3% (33/53), $1.1{\times}10^8{\pm}2.8{\times}10^8IU/mL$ 3) Group III (IRMA negative, CMIA negative, N = 36), HBV DNA detection rate 36.1% (13/36), $3.0{\times}10^5{\pm}1.1{\times}10^6IU/mL$ 4) Group IV(IRMA positive, CMIA negative, N = 12), HBV DNA detection rate 25% (3/12), $1.3{\times}10^3{\pm}1.1{\times}10^3IU/mL$ Conclusion HBeAg detection rate according to the serological test showed a large difference. This difference is considered for a number of reasons such as characteristics of the Ab used for assay kit and epitope, HBV of genotype. Detection rate and the concentration of the group-specific HBV DNA classified serologic results confirmed the high detection rate and the concentration in Group II (IRMA-positive, CMIA positive, N = 53).

HBV-trimera 동물모델을 통한 B형 간염 항체의 효능평가에 관한 연구

강영국(Young Kook Kang),장명희(Myeong Hee Jang),김근수(Keun-Soo Kim),오미숙(Mee Sook Oh),김남재(Nam Jae Kim),이병석(Byung Seok Lee),이은나(Eun Na Lee),김성주(Sung Joo Kim),류춘제(Chun Jeih Ryu),홍효정(Hyo Jeong Hong) 한국실험동물학회 2005 Laboratory Animal Research Vol.21 No.1

Humanized Balb/c mice (termed Trimera mice) conditioned by lethal total body irradiation and bone marrow transplantation from NOD/SCID mice have been described to support efficient engraftment of human tissues. To evaluate the efficacy of potential anti-HBV agents in vivo, we, here, describe the development of a mouse Trimera model for human hepatitis B virus (HBV) infection. HBV viremia was induced by transplantation of ex-vivo HBV-infected human liver fragments under the kidney capsule of Trimera mice (HBV-Trimera). The levels of HBV viremia were determined by measuring serum HBV DNA using polymerase chain reaction (PCR) at 10 days after liver transplantation. In order to evaluate the therapeutic potential of two humanized monoclonal antibodies specific to hepatitis B surface antigens, the HBV-Trimera mice were administrated intraperitoneally with anti-HBs antibody (HzSIII) or anti-preS1 antibody (AP301) at days 14 to 17 post-liver transplantation. Treatment of the HBV-Trimera mice with two anti-HBV humanized monoclonal antibodies (HzSIII, AP301) reduced the viral load in their sera in a dose-dependent manner, suggesting that the humanized antibodies will be useful in the prevention and treatment of HBV infection. These results suggest that the HBV-Trimera mice can be used as an animal models for evaluating therapeutic efficacy of anti-HBV agents.

B형 간염바이러스의 태아와 성인간세포에 대한 감염성 해명 연구

정태호,김성국,서화정,윤영국,임종완,차상호 경북대학교 의학연구소 1999 경북대학교병원의학연구소논문집 Vol.3 No.1

사람의 태아와 성인의 간세포에 대한 B형 간염바이러스의 감염도를 비교 분석함으로써 HBV 와 간세포간의 친화도내지는 HBV 수용체에 관한 정보를 얻고자 함 대상 및 방법 : 성인이 외상을 입어 부득이 손상된 간조직을 절제하였을 경우와 모자보건법상 임신지속이 어려운 상태에서 부인과적 수술시 21주 전후의 태아 간조직을 얻어 DMEM 을 배지로 50% 용량으로 HBsAg 양성인 사람 혈청을 가하여 5일간 체외 배양하고 병리조직학적 처리후 효소 면역학적 방법으로 HBsAg 를 염색하여 바이러스의 존재 여부를 검색하였다. 결과 : 태아와 성인 간조직을 5일간 체외 배양하였을 때 그 조직상은 간조직임을 인지할 수 있었다. HBV 의 감염여부는 DAKO 사의 효소면역 법으로 조사한 결과 태아의 간조직에서는 HBsAg 양성부위를 발견하지 못하였으나, 성인 간조직에서는 HBsAg 양성부위가 발견되었다. 결론 : 태생 21주 전후의 태아의 간세포는 HBV 와 친화도는 아주 미약하거나 없고, 성인 간세포는 HBV에 대한 친화도가 높다. 따라서 태생 21주의 간세포에는 HBV 의 감염이 어렵다는 것을 인지하게 되었다. Considerable progress has been made in the immune prophylaxis of hepatitis B virus(HBV) infection and in the understanding of its replication and pathogenecity at the molecular level. However, a convenient in vitro assay for HBV infectivity has been lacking and the early steps of viral life cycle are now yet well understood. Primary cultured human hepatocytes are reported to be susceptible to HBV 10-12, but these systems are not practical because they are short-lived and require primary explanted human liver. However, it is uncertain yet whether primary hepatocytes of human fetal liver are susceptible to HBV for infection or not. We have expanded the infectivity of HBV into a primary cultures of human fetal hepatocytes together with controversy of adult primary hepatocytes. To investigate the infectivity of HBV for, adult and fetal hepatocytes After 5 day culturing fetal(1) and adult(2) liver tissues, which obtained from a surgical operation on DMEM culture media with half HBV positive serum, separating liver cells and fixing them, the existence of HBsAg antigen was investigated through the immuno-enzymatic staining. HBsAg stained area was not found in the 21 weeks gestation liver cells until 5 days of invitro-culture, while it was found in the adult liver cells. This test result showed that the HBV infection is very hard to happen in the liver cells 21 weeks gestation or prior one.

Critical role of bile acid (BA) in the cellular entry and permissiveness of Hepatitis B virus in vitro

Hong‑Jai Lee,Bo‑Young Shin,Jae‑Seung Moon,Ailyn Fadriquela,Selikem Abla Nuwormegbe,Chun‑Chang Ho,Jin‑Su Shin,Jee‑Sang Yoon,Sang‑Kyou Lee,Soo‑Ki Kim 대한독성 유전단백체 학회 2020 Molecular & cellular toxicology Vol.16 No.3

Background Lack of universal replication system for hepatitis B virus with narrow host range and organ tropism has hampered to uncover the pathogenesis of HBV. Previously, we reported the essentiality of humoral milieu and its components toward HBV and hepatitis C virus survival/viability in vitro. Of these components, the precise role of enterohepatic humoral milieu such as bile acid (BA) on HBV cultivation in vitro and in vivo is unknown. Objective We explored whether BA, specifically taurochenodeoxycholic acid (tCDCA) would directly regulate the viral DNA and surface antigen expression of HBV in vitro, consequently rendering HBV to enter into human or murine immortalized hepatocytes, and non-hepatocytes. Result We found that higher concentration of taurochenodeoxycholic acid (tCDCA) is able to preserve the genomic stability of HBV in cell-free DMEM, showing higher the surface antigenicity than taurocholic acid (tCA). In line, we found that in vitro cell culture condition (100 μmol/L of tCDCA coupled with 1 × 108 g e/mL HBV) would be optimal for HBV entry into target cells. Using this, human (HepG2, Huh7), and rodent (Hepa1c1c7, H4-II-E) hepatoma cell lines were infected by HBV, as evidenced by the presence of HBV biomarkers (HBsAg, and HBV DNA in culture supernatant, as well as HBcAg in cell). Further, cellular entry test revealed that HBV is able to infect 12 different non-hepatic cell lines regardless of species, and organ/tissue, consequently reproducing progeny as confirmed by HBV biomarkers. Last, reinfection test showed that the progenies of HBV from immortalized HepG2, and Hepa1c1c7 cells are able to enter into each or vice versa naïve HepG2, and Hepa1c1c7 cells with or without BA. Conclusion This study demonstrates that enterohepatic humoral milieu such as BA, specifically tCDCA would directly regulate HBV DNA and its surface antigen expression in vitro, consequently rendering HBV to enter into human or murine immortalized hepatocytes, and non-hepatocytes. This is the first note to render HBV permissive to human or rodent hepatic and non-hepatic cells via sole manipulation of humoral milieu, thus establishing the platform for in vitro robust replication system of HBV.

Comparative Study of Persistent Immunity to HBV after Vaccination and Naturally Acquired Immunity Post HBV Infection in Mongolia

( O. Otgonbayar ),( Dashchirev Munkh-orshikh ),( Oidov Baatarkhuu ) 대한간학회 2020 춘·추계 학술대회 (KASL) Vol.2020 No.1

Aims: To determine the generation of persistent immunity from HBV vaccination or after HBV infection. HBV and HCV are high prevalent in Mongolia, around one fourth Mongolian has HCV or HBV. According to the study, 39.4% of people who had HBV infection got HBV vaccination which leads to viral problem in Mongolian health care system. Since 1991 all newborns have been vaccinated with HBV vaccine. And people who born before that year did not take the HBV vaccination. Methods: 492 patients have enrolled who were investigated with quantitative HBsAb using Sysmex HISCL-800 at Happy Veritas Clinic and Diagnostic Center and MNUMS. The vaccination scheme consists of three doses. Vaccination is successful if the antibody titer is higher than 10 mlU/L. Also we have conducted questionnaires about HBV vaccination and risk factor for taking hepatitis infections from patients. Results: In this study 492 patients have participated 313(63%) female and 179 (37%) male, out of which 471(96%) people born before 1991 and remaining 21(4%), people born after 1991. 12 people (57%) who born after 1991 or vaccinated within 24 hours after birth were quantification HBsAb low titer (< 10mlU/L), remaining (43%) were qHBsAb titer ( >10mlU/L), while 297 people (64%) who born before 1991 were qHBsAb titer (<10mlU/L),and remaining 36% of patients had persistent HBV vaccine. The 99 people who born before 1991 have enrolled in HBV vaccination voluntarily while 372 people did not take HBV vaccine at all. Conclusions: Persistent immunity against HBV is generated not only in person who have taken HBV vaccination but also in person who have had slight HBV infection. It was considered that people aged between 50 and 60 years could not get persistent immunity against HBV. We assumed that persistent immunity against HBV depends on age, not other factor and sex.