Turkish Journal of Medical Sciences

Detection of Hepatitis Virus Surface Antigen (HBsAg) with Avidin/Biotin Solid-Phase Enzymeimmunoassay (EIA)




An avidin/biotin solid-phase sandwich enzymeimmunoassay for the detection of hepatitis B virus surface antigen (HBsAg) was set up. Hepatitis B virus surface antigen was used for immunization of rabbits. Resulting polyclonal antibodies (Ig fraction) were precipitated by the salting-out procedure (Na_2SO_4) and biotinylated using a succinimide ester of biotin. Streptavidin-conjugated horseradish peroxidase (HRP) was used as a probe detector reagent. The assay was compared with two different commercial HBsAg detection ELISA kits utilizing monoclonal antibodies to HBsAg (Wellcozyme and Sorin), using pre-screened 82 positive and 67 negative serum samples. Despite the fact that three false negative results were obtained our assay was comparable to the other methods in detecting negative and positive samples. Although 3.66 % of the results were false negative, this preliminary work may offer a new HBsAg screening method for country and our reference preparation of HBsAg detection EIA kit can be substitued for expensive imported commercial kits.


HBsAg Detection, Avidin-biotin System, ELISA

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