Development of sandwich ELISA systems for the diagnosis of hepatitis B virus surface antigen and its antibody in human sera

Abstract

Hepatitis B is an infection of the liver caused by the Hepatitis B virus HBV HBV is one of the major causes of acute and chronic hepatitis cirrhosis and hepatocellular carcinoma and it is a serious global public health problem It is estimated that million individuals worldwide are infected with the virus which causes deaths each year People most at risk include antibody who has unprotected sexual intercourse drug users who share needles and syringes health care workers in contact with potentially contaminated blood or body fluids anyone in intimate contact with the infected person Therefore the development of economic and accurate detection systems and potential alternative antiviral approaches for HBV detection could be important for fighting viral hepatitis Hepatitis B diagnosis has been based on the detection of serologic markers Testing for these markers helps to determine the presence of past or ongoing HBV infection the acute chronic or subclinical carrier state of the disease response to therapy and or the immune status of the patient Hepatitis B virus surface antigen HBsAg is the first serological marker to appear in the circulation well before clinical symptoms and is the viral component usually found in the highest concentration in the serum of HBV infected patients The presence of anti HBs in serum indicates previous exposure to HBV and long lasting acquired immunity Low serum titres of anti HBs however it can signal a lack of immunity to future HBV infection In this study diagnostic systems based on the use of monoclonal antibody MAb and polyclonal antibodies PAb were developed and conjugated enzyme and biotin for early and sensitive diagnosis of Hepatitis B which is still one of the important infectious diseases in Turkey Sandwich ELISA kit systems were generated by using both G MAb as capturing agent and G HRP or G biotin conjugates as detecting agents Homemade sandwich ELISA tests were compared with the other conventional sandwich ELISA tests by using hepatitis B positive and negative infection serum They were shown that our system gave reliable results When the homemade HBsAg ELISA system were compared with the other commercial kit by using patients sera it was shown that our system corresponded with the results of negative and positive samples at ratio of When the homemade anti HBsAg ELISA system was compared with the commercial kit by using patients sera it was shown that our system corresponded with the results of negative and positive samples at ratio of In subsequent studies by the final tests of homemade ELISA kit it was observed that Biotin labeled kits responded very close results with the conformity level when compared with commercial kits