HBsAg ELISA KIT
INTENDED USE
This HBsAg ELISA Kit is to be used for the in vitro detection of Hepatitis B surface antigen (HBsAg) including the subclass of HBsAg (ad, ay.) in human plasma or serum. This HBsAg ELISA Kit is intended for RESEARCH USE ONLY, and is not to be used in diagnostic or therapeutic procedures.
INTRODUCTION
Hepatitis B is an infection of the liver caused by the hepatitis B virus. Approximately 5-10% of adults and 90% of babies who are infected with HBV will go on to carry the virus for the rest of their lives. These people will pass the virus onto others. HBV is excreted in body fluids such as semen, saliva, blood and urine in persons with acute or chronic infection. The route of transmission can include homosexual or heterosexual activity, blood-borne exposure (needles, transfusion), mother-infant, close personal contact and even by consuming contaminated food or water. Thus, Hepatitis B has become a major public health concern. When HBV invades the body it causes liver damage through induction of auto-immunity. Liver cell injury results from cytotoxic T cell activity rather than viral cytotoxic activity. The principle screening test for detecting current (acute or chronic) HBV infection is the identification of HBsAg, an envelope lipoprotein. This is the first immunological marker, to appear in a patient's serum and exist in high quantities in the blood. Patients who are HBsAg positive develop chronic persistent hepatitis (CPH) and chronic active hepatitis (CAH). Patients with CPH usually remain in good health but those with CAH have progressive liver damage with the outcome being portal fibrosis, cirrhosis, or hepatocellular carcinoma.
Screening for HBsAg is recommended for all donors, pregnant women and those individuals who at high risk. Presence of this marker is conclusive proof of HBV infection.
PRINCIPLE OF THE ASSAY
HBsAg ELISA Kit
HBsAg ELISA Kit is to be used for the in vitro detection of Hepatitis B surface antigen (HBsAg) including the subclass of HBsAg (ad, ay.) in human plasma or serum. This HBsAg ELISA Kit is intended for RESEARCH USE ONLY, and is not to be used in diagnostic or therapeutic procedures.
Monoclonal antibodies specific for HBsAg have been bound to the surface of each microplate well. During the course of the assay, the positive control, negative control and samples are added to the microplate wells. The microtiter plate wells are thoroughly washed to remove unbound other components of the sample. A standardized preparation of horseradish peroxidase (HRP) conjugated antibody specific for HBsAg is added to each well to “sandwich” the antibodies immobilized during the first incubation. Following a wash to remove any unbound HRP conjugate, a TMB (3.3’, 5,5' tetramethyl-benzidine) substrate solution is added to each well. The enzyme (HRP) and substrate are allowed to react over a 10-minute incubation period. The enzymesubstrate reaction is terminated by the addition of sulfuric acid solution and the color change is measured spectrophotemetrically at a wavelength of 450nm±2nm. Only those wells containing HBsAg and HRP conjugate will exhibit a change in color. The intensity of this color change is proportional to the concentration of HBsAg in the sample.
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