KSBB Journal

The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
권호 표지

Development of Diagnostic kit for Hepatitis B Susrface Antigen using Immunochromatographic Assay Method

면역크로마토그래피법을 이용한 B형간염 진단용 kit의 개발

  • Published : 2000.04.01
Download PDF
⟨ Previous Next ⟩

Abstract

A hepatitis B Surface Antigen(HBsAg)-screening kit using immunochromatographic assay(ICA) method was developed by e employing two kinds of antibodies. One is mouse monoclonal anti-HBs for tracer antibody and the other is goat p이yclonal a anti-HBs for capture antibody. This capture antibody was immobilized on the surface of nitroceliulose(NC) membrane and the t tracer antibody was conjugated with g미d particles. When serum sample was added to the sample well, the $\infty$njugates d deposited in a dry state on the surface of glass fiber filter were reconstituted and then combined with HBsAg in serum. In 5 5 min after adding, the assay result was visible through the window, that is, the complexes composed of HBsAg and the c conjugates appeared as maroon line on the lower part of the NC membrane. The detection limit of the ICA kit was 2 ng/ml w when being tested with the reference HBsAg.

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.

Keywords

References

  1. Scand. J. Infect. Dis. v.7 no.2 Some epidemiological and clinical aspects of hepatitis Bantigen and its subtypes Nordenfelt, E.
  2. Jpn. J. Exp. Med. v.60 no.5 Common and subtypic determinants of hepatitis B surface antigen particles: susceptibility to reduction and/or alkylation evaluated with monoclonal antibodies Luengrojanakul, P.;H. Ohnuma;K. Tachibana;S. Usuda;H. Okamoto;T. Tanaka;F. Tsuda;A. Machida;M. Mayumi
  3. J. Virol. Methods v.33 no.1-2 Determination of HBsAg subtypes in different high risk populations using monoclonal antibodies Swenson, P.D.;J.T. Riess;L.E. Krueger
  4. Dev. Biol. Stand. v.30 Quantitative derection and typing of HBsAg and anti-HBs by radioimmunoassay techniques Petrilli, F.L.;P. Crovari;S. De Flora
  5. J. Clin. Pathol. v.29 A comparative study of passive haemagglutination methods for the detection of hepatitis B suface antugen in toutine hospital practice Withers, M.J.;G.V. McCahill;P.D. Griffiths;R.B. Heath;Jr Pattison;D.S. Dane
  6. J. Virol. Methods v.2 Rapid detection of HBsAg and anti-HBs by enzyme-immunoassay Haas, J.
  7. J. Med. Microbiol. v.16 Comparison of enzyme immunoassay with radioimunoassay for the detection of hepatitis B surface antigen Hendry, R.M.;L.W. Herzog;J.E. Herman
  8. J. Histochem. Cytochem. v.25 Absorption of horse radish peroxidase, ovomucoid and anti-immunoglobulin to colloidal gold for the indirect detection of concanavalin A, wheat germ agglutinin and goat anti-human immunoglobulin G on cell surfaces at the electron microscope level A new method, theory and application Geoghegan, W.D.;G.A. Ackerman
  9. Proc. Natl. Acad Sci. v.78 Colloidal gold-low density lipoprotein conjugates as membrane receptor brobes Chien, S.
  10. World patent application WO 88/08534 May, K.;M.E. Prior;I. Richards
  11. J. Clin. Microbiol. v.35 Serodiagnosis of Helicobacter pylori infection: comparison and correlation between enzyme-linked immunsorbent assay and rapid serological test results Chen, T.S.;F.Y. Chang;S.D. Lee
  12. Tuber. Lung Dis. v.77 Chinical evaluation of a rapid immunochromatiographic assay based on the 38 kDa antigen of Mycobacterium tuberculosis on patients with pulmonary tuberculosis in China Cole, R.A.;H.M. Lu;Y.Z. Shi;J. Wang;De-Hua T.;A.T. Zhou
  13. Vet. Rec. v.138 Evaluation of an one-step test for rapid, in pratice detection of rotavirus on farm animals Klingenberg, D.V.K.;Espandiari, J.
  14. Acta. Chem. Scand. v.50 Immunochromatographic assay for quantitation of milk progesterone Laitinen, M.P.;Vuento, M.
  15. J. Clin. Microb. v.36 Evaluation of a rapid immunochromatographic test for diagnosis of Dengue virus infection Vaughn, D.W.;A. Nisalak;S. Kalayanarooj;T. Solomon;N.M. Dung;A. Cuzzubbo;P.L. Devine
  16. J. Clin. Microbiol. v.34 Evaluation of immunochromatographic assay systems for rapid detection of hapatitis B surface antigen and antibody, Dainascreen HBsAg and Dainascreen Ausab Kumiko, S.;I. Satoshi;I. Yoshitsugu;N. Toshi;S. Kaoru;N. Nobuo
  17. Nature Physical Science v.241 Controlled nucleation for the regulation of the partucle size in monodisperse suspensions Frens, G.
  18. Electrophoresis v.19 Evidence of intra - and extracellular modifications of monoclonal IgG polypeptide chains generating charge heterogeneity Mimura, Y.;K. Nakamura;T. Tanaka;M. Fujimoto
  19. J. Histochem. cytochem. v.25 Colloidal gold, a useful marker for transmission and scanning electron microscopy Horisberger, M.;J. Rosset
  20. J. Histochem. Cytochem. v.36 The effect of three variables on adsorption of rabbit IgG to colloidal gold Geoghegan, W.D.
  21. Adv. Colloid Interface Sci. v.25 Adsorption of proteins from solution at the solid-liquid interface Norde, W.