• 제목/요약/키워드: hepatitis B surface antigen(HBsAg)

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Expression of Hepatitis B Virus S Gene in Pichia pastoris and Application of the Product for Detection of Anti-HBs Antibody

  • Hu, Bo;Liang, Minjian;Hong, Guoqiang;Li, Zhaoxia;Zhu, Zhenyu;Li, Lin
    • BMB Reports
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    • 제38권6호
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    • pp.683-689
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    • 2005
  • Antibody to hepatitis B surface antigen (HBsAb) is the important serological marker of the hepatitis B virus (HBV) infection. Conventionally, the hepatitis B surface antigen (HBsAg) obtained from the plasma of HBV carriers is used as the diagnostic antigen for detection of HBsAb. This blood-origin antigen has some disadvantages involved in high cost, over-elaborate preparation, risk of infection, et al. In an attempt to explore the suitable recombinant HBsAg for the diagnostic purpose, the HBV S gene was expressed in Pichia pastoris and the product was applied for detection of HBsAb. Hepatitis B virus S gene was inserted into the yeast vector and the expressed product was analyzed by sodium dodecyl sulphate polyacrolamide gel electrophoresis (SDS-PAGE), immunoblot, electronic microscope and enzyme linked immunosorbent assay (ELISA). The preparations of synthesized S protein were applied to detect HBsAb by sandwich ELISA. The S gene encoding the 226 amino acid of HBsAg carrying ahexa-histidine tag at C terminus was successfully expressed in Pichia pastoris. The His-Tagged S protein in this strain was expressed at a level of about 14.5% of total cell protein. Immunoblot showed the recombinant HBsAg recognized by monoclonal HBsAb and there was no cross reaction between all proteins from the host and normal sera. HBsAb detection indicated that the sensitivity reached 10 mIu (micro international unit)/ml and the specificity was 100% with HBsAb standard of National Center for Clinical Laboratories. A total of 293 random sera were assayed using recombinant S protein and a commercial HBsAb ELISA kit (produced by blood-origin HBsAg), 35 HBsAb positive sera and 258 HBsAb negative sera were examined. The same results were obtained with two different reagents and there was no significant difference in the value of S/CO between the two reagents. The recombinant HBV S protein with good immunoreactivity and specificity was successfully expressed in Pichia pastoris. The reagent for HBsAb detection prepared by Pichia pastoris-derived S protein showed high sensitivity and specificity for detection of HBsAb standard. And a good correlation was obtained between the reagent produced by recombinant S protein and commercial kit produced by blood-origin HBsAg in random samples.

B형간염바이러스 및 C형간염바이러스의 표식자 양성율과 원발성 간세포 암의 연관성에 대한 환자-대조군 연구 (A Case-Control Study on Association Between Hepatocellular Carcinoma and Infection of Hepatitis B and Hepatitis C Virus)

  • 안형식;김민호;김영식;김정순
    • Journal of Preventive Medicine and Public Health
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    • 제30권1호
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    • pp.1-15
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    • 1997
  • To investigate the association between hepatocellular carcinema(HCC) and infection of hepatitis B virus(HBV) and hepatitis C virus(HCV) in an HBV endemic area, a case-control study of 254 patients with HCC and of 1,270 age and sex matched health control subjects was done. Among the 254 HCC patients 166(65.4%) were positive for hepatitis B surface antigen(HBsAg), 49(19.3%) were positive for HCV antibody (anti-HCV Ab). The crude odd ratio of patients with HBsAg was 36.1(95% CI :22.4-58.2) and with anti-HCV Ab was 9.0(95% CI :5.5-14.6). In an analysis, which HBsAg(-), HBcAb(-), anti-HCV Ab(-) group was chosen as referent group, odd ratio of HBsAg(+) group was 14.4(95% CI: 7.2-28.9) and of anti- HCV Ab(+) was 10.7(95% CI: 2.9-40.0). odd ratio of anti-HCV Ab(+), HBsAg(+) group and anti-HCV Ab(+), HBsAg(-), HbcAb(+) group for HCC were elevated to 27.3(95% CI : 9.0-82.9), 15.9(95% CI:7.1-35.8) respectly, The odd ratio of anti-HCV Ab(-), HBsAg(-), HBcAb(+) group was 2.4(95% CI : 1.1-5.0). These result suggested that HBV and HCV were associated with HCC. In HBV endemic area patients with HBcAb alone should be considered risk group for HCC.

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면역크로마토그래피법을 이용한 B형간염 진단용 kit의 개발 (Development of Diagnostic kit for Hepatitis B Susrface Antigen using Immunochromatographic Assay Method)

  • 신형순;신광순;정홍근;허태련
    • KSBB Journal
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    • 제15권2호
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    • pp.214-218
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    • 2000
  • 면역크로마토그래피 진단방법을 이용하는 B형간염 스크리닝 kit를 개발하기 위하여 두가지의 항체를 이용하였다. 표식자항체로 사용된 것은 단세포군항체 anti-HBs이고 포획항체는 goat anti-HBs 인데 포획항체는 니트로셀룰로즈 막에 고정되고 표식자항체는 금 입자에 결합된다. 혈청 검체를 well에 가하면 유리섬유 표면에 건조상태로 침착되어 있던 conjugate가 활성화되어 검체중의 HBsAg와 결합한다. 검체를 가한 지 5분 후 검사결과가 나타나는데 HBsAg와 conjugate가 결합된 복합체가 니트로셀롤로즈 막의 하단부에 붉은 색 선으로 나타난다. 본 kit의 검출한계는 표준 HBs-Ag 용액을 사용하여 시험하였을 때 2 ng/ml이었다.

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B형 간염 바이러스 양성인 만성 간질환에서 Hepatitis B 표면항원의 아형 (Subtypes of Hepatitis B Surface Antigen Among Chronic Liver Disease)

  • 조희순;임소여;이채훈;김경동;김정숙
    • Journal of Yeungnam Medical Science
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    • 제13권2호
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    • pp.272-278
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    • 1996
  • B형 간염 표면항원(HBsAg)의 4가지 주요 아형 조사를 통해 HBsAg과 B형 간염표면항체가 모두 양성인 환자에서 항원과 항체의 아형이 다름을 밝힘으로 항원-항체 공존을 이해하게 되었으며, 감염경로를 추적하는 역학적 조사, 인구이동양상의 지표에 유용하게 사용될 수 있다. 현재까지 우리나라의 HBV 아형에 관한 연구는 전체적인 빈도의 조사나 HBsAg과 anti-HBs가 공존하는 경우에서 아형이 가지는 의미에 대한 것이 전부였다. 본 연구에서는 영남대학교 의과대학 부속병원에서 만성 간질환으로 진단받고 경과추적중인 HBsAg 양성인 환자 214명을 대상으로 혈청에 존재하는 HBsAg 아형을 단세포군 항체를 이용한 혈청학적 방법으로 조사하였다. 214명의 검체중 206명(93.9%)에서 adr, 6명(2.8%)에서 adw, 2명(0.9%)에서 ayr, 2명(0.9%)에서 ar, 3명(1.4%)에서 adwr로 나타났으며 ayw는 한 례에서도 발견되지 않았다. 아형과 질환의 중증도 사이에는 유의한 연관성은 발견되지 않았다. 결론적으로 만성 간질환 환자에서 adr이 가장 많고 질환의 중증도와 아형간에 유의한 연관성은 발견되지 않았지만, 아형이 만성 B형 바이러스 간염의 간경변증이나 간세포암으로의 진행과 예후에 영향을 미치는지 밝히기 위해서는 좀 더 연구를 진행하여야 할 것이다.

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연잎추출물의 B형 간염 표면 항원 발현 억제 효과 (Nelumbo nucifera Leaves Extract Reduced the Production of Hepatitis B Surface Antigen on HepG2.2.15)

  • 이윤희;강이중;이성진
    • 한국약용작물학회지
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    • 제17권2호
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    • pp.133-138
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    • 2009
  • Nelumbo nucifera (lotus) is known to be a useful medicinal plant and leaf extract contains several flavonoids and alkaloids. To analyze the effect of Nelumbo nucifera leaves extract (NNL) on the HBsAg production, we treated NNL on HepG2.2.15 cells which contain the hepatitis B viral genome and secrete surface antigen into media. NNL suppressed the production of hepatitis B surface antigen as a dose-dependent manner. To analyze the effect of NNL on HBV DNA replication, PCR analysis was performed. NNL was not affected the HBV DNA replication and HBsAg mRNA expression. To understand the effect of NNL on the production of HBsAg, we carried out the analysis of lipid-metabolizing gene expression using one-step RT-PCR. NNL reduced the gene expression of FASN and SREBP2 and increased the expression of LDLR. Triglyceride content of HepG2.2.15 cells was not decreased by treatment of NNL. This result suggests a possibility that NNL may have an effect for the inhibition of hepatitis B surface antigen by modulation of lipid and cholesterol metabolism.

군 입영자에서의 B형 간염 발생 규모와 B형 간염 바이러스 보유자가 이에 미치는 영향 (The Incidence of Hepatitis B in Military Service and the Effect of Asymptomatic HBsAg Carriers on the Incidence)

  • 김록권;서일;남정모;한광협
    • Journal of Preventive Medicine and Public Health
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    • 제30권2호
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    • pp.267-278
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    • 1997
  • The purpose of this study was to investigate the incidence rate of hepatitis B in the military service and to examine the effect of the asymptomatic hepatitis B surface antigen(HBsAG) carries on the incidence of hepatitis B. The subject were 223,270 men who were conscripted to the Korean Army from 1991 to 1994. According to the conscripted year, four conscription cohort were constructed. At the screening examination for military service no test for hepatitis B were performed in 1991 and 1992. In 1993, a screening test for hepatitis B were performed and those who were confirmed as HBsAg positive o. showed high titers $(\geqq100IU)$ of nm glutamic-pyruvic transaminase(SGPT) were excluded from conscription. In 1994, the criteria for conscription was changed and those who were HBsAg positive were not excluded from conscription. Only those who showed $\geq$ SGPT 100IU were excluded. The main results were as follows ; 1. The positive rate of HBsAg is 5.5% in the conscripted men. 2. The incidence rates of the hepatitis B in 1991 and 1992 conscription cohort were 9.96 and 8.10 per ten thousand person-year, respectively. The incidence rate of the hepatitis B was 1.34 per ten thousand person-year in 1993 conscription cohort which was confirmed as HBsAg negative at the screning test, and 7.41 per ten thousand person-year in 1994 conscription cohort which included the HBsAg positive. 3. The incidence rate of hepatitis B was 99.98 per ten thousand person-year in HBsAg positive group and 2.25 per ten thousand person-year in HBsAg negative group. The incidence rate of the group with high SGPT and HBsAg positive was 255 times higher than that of normal population. 4. The incidence of hepatitis B in HBsAg negative group did not increase even though the probability of personal contact with HBsAg positive had been increased. from the above results, the men who have high SGPT with HBsAg positive should be excluded from military service, and it can not be said that asymptomatic HBsAg carriers influence on the hepatitis B incidence among the HBsAg negative through personal contact.

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유전자 재조합 B형 간염 바이러스 표면 항원, CJC-50100의 일반약리작용 (General Pharmacology of CJC-50100, a Recombinant Hepatitis B Virus Surface Antigen)

  • 정성학;최재묵;이남중;전형수;김연희;김재승;하석훈;김영훈;이나경
    • Biomolecules & Therapeutics
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    • 제9권1호
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    • pp.55-62
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    • 2001
  • CJC-50100 is a recombinant hepatitis B virus surface antigen (HBsAg) expressed in yeast. The general pharmacological properties of CJC-50100 were evaluated in mice, rats, dogs and isolated guinea pig ileum. The doses were 0.33~33.3 $\mu$g/kg i.m. for mice and rats and 3.3~9.9 $\mu$g/kg i.v. for dogs. The concentrations of 0.002~0.02 $\mu$g/ml were used for the assay with guinea pig ileum. Intramuscular administration of CJC-50100 at the doses did not alter general behavior and the responses for central nervous system, smooth muscle, gastrointestinal system, cardiovascular and respiratory system, and water and electrolytes excretion. In summary, CJC-50100 had no pharmacological effect in these studies even up to the 100-fold of the expected clinical dose, 20 $\mu$g/man/60 kg.

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치과대학생 및 전공 에 있어서 B형간염면항원의 발현빈도에 관한 연구 (A Study on the Occurence Rate of HBsAg in a Dental School Population)

  • 이건복;정성창
    • Journal of Oral Medicine and Pain
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    • 제7권1호
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    • pp.66-70
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    • 1982
  • Medical personnek are one of several groups that have been reported to have a high incidence of hepatiris B. It is also thought that the occurrence rate of hepatitis B surface antigen(HBsAg), aserologic marker for hepatitis B virus(HBV), is expected to be high in the dental personnel who are frequently exposed to the blood and saliva of the patients. Although many studies have been done to determine the HBsAg status of virus groups, limited investigations have been performed on dental personnel, especially in this country. The main purpose of this study was to identify HBsAg positivity among dental students, interns, and residents who would be expected to be a high risk group of hepatitis B infection. Screening test for HBsAg of a dental school population was performed by indirect hemagglutination(IHA)in 1982. The results were as follows : 1. Thirty four out of a total 362 persons(9.4%)tested in the study had positive response for HBsAg in their serum samples. 2. Twenty seven out of 320 dental students(8.4%)had positive for HBsAg, and in Senior class 12 out of 82 dental students (14.6%)had positive response that was the highest incidence among dental students group. 3. Seven out of 42 interns and residents(16.7%)had positive for HBsAg, and it was the highest incidence in this dental school population.

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Development of a Rapid Automated Fluorescent Lateral Flow Immunoassay to Detect Hepatitis B Surface Antigen (HBsAg), Antibody to HBsAg, and Antibody to Hepatitis C

  • Ryu, Ji Hyeong;Kwon, Minsuk;Moon, Joung-Dae;Hwang, Min-Woong;Lee, Jeong-Min;Park, Ki-Hyun;Yun, So Jeong;Bae, Hyun Jin;Choi, Aeran;Lee, Hyeyoung;Jung, Bongsu;Jeong, Juhee;Han, Kyungja;Kim, Yonggoo;Oh, Eun-Jee
    • Annals of Laboratory Medicine
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    • 제38권6호
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    • pp.578-584
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    • 2018
  • Background: Accurate, rapid, and cost-effective screening tests for hepatitis B virus (HBV) and hepatitis C virus (HCV) infection may be useful in laboratories that cannot afford automated chemiluminescent immunoassays (CLIAs). We evaluated the diagnostic performance of a novel rapid automated fluorescent lateral flow immunoassay (LFIA). Methods: A fluorescent LFIA using a small bench-top fluorescence reader, Automated Fluorescent Immunoassay System (AFIAS; Boditech Med Inc., Chuncheon, Korea), was developed for qualitative detection of hepatitis B surface antigen (HBsAg), antibody to HBsAg (anti-HBs), and antibody to HCV (anti-HCV) within 20 minutes. We compared the diagnostic performance of AFIAS with that of automated CLIAs-Elecsys (Roche Diagnostics GmbH, Penzberg, Germany) and ARCHITECT (Abbott Laboratories, Abbott Park, IL, USA)-using 20 seroconversion panels and 3,500 clinical serum samples. Results: Evaluation with the seroconversion panels demonstrated that AFIAS had adequate sensitivity for HBsAg and anti-HCV detection. From the clinical samples, AFIAS sensitivity and specificity were 99.8% and 99.3% for the HBsAg test, 100.0% and 100.0% for the anti-HBs test, and 98.8% and 99.1% for the anti-HCV test, respectively. Its agreement rates with the Elecsys HBsAg, anti-HBs, and anti-HCV detection assays were 99.4%, 100.0%, and 99.0%, respectively. AFIAS detected all samples with HBsAg genotypes A-F and H and anti-HCV genotypes 1, 1a, 1b, 2a, 2b, 4, and 6. Cross-reactivity with other infections was not observed. Conclusions: The AFIAS HBsAg, anti-HBs, and anti-HCV tests demonstrated diagnostic performance equivalent to current automated CLIAs. AFIAS could be used for a large-scale HBV or HCV screening in low-resource laboratories or low-to middle-income areas.

A Study on the Positive Rates of HBsAg and Anti-HBs from Old People's Home and Casual Ward in Daegu, Korea

  • Shin, Hyun Gyu
    • 대한임상검사과학회지
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    • 제43권4호
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    • pp.145-149
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    • 2011
  • Serum hepatitis is still recognized as a major public health problem in many countries. The most common etiologies are acute viral hepatitis A and B, and hepatitis B virus (HBV) was known as the pathogen of serum hepatitis. since the HBV causes chronic hepatitis. HBV infection is hyperendemic in Korea. It is known that 8% and 1% of the Korean population is chronic carriers of HBV. The total number of 487 serum specimen (old people's home 190, causal ward-Daegu hope village 297) were collected at the laboratory of Department of Clinical Pathology of Daegu medical center. The laboratory tested the specimen to detect for hepatitis B surface antigen (HBsAg) and anti-HBs using reverse passive hemag-glutination (RPHA) and passive hemagglutination (PHA) method between April, 2011 and June, 2011. In one-step test, HBsAg and anti-HBs of Humasis were used. The positive rate for HBsAg was 6.80% (33/487), and the rate in male was 6.93% (19/274) higher than that of female 6.57% (14/213). In the positive rate of HBsAg by age group, the highest positive rate group was in the group of 40-49 years in male (12.0%), and 50-59 years in female (17.65 %). The overall positive rate for anti-HBs was 42.70% (208/487), showing the higher positive rate of 43.80% in male (120/274) than that of 41.30% (88/213) in female. The highest positive rate of anti-HBs in age group was the 40-49 years group in male (52.00%) and the group of under 29 in female (66.67%).

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