HBsAg, or Hepatitis B surface Antigen, is a protein present on the surface of the Hepatitis B virus. Its presence in the blood indicates an ongoing Hepatitis B infection, making it a crucial marker for diagnosis.
The HBsAg ELISA test is a laboratory method used to detect the presence of HBsAg in a patient's serum or plasma. ELISA stands for Enzyme-Linked Immunosorbent Assay, and it provides a sensitive and specific way to diagnose Hepatitis B.
The first step is collecting a blood sample from the patient. This is typically done via venipuncture, where a needle is used to draw blood from a vein, usually in the arm.
After collection, the blood sample is processed to separate the serum or plasma. This is done by centrifuging the blood, which separates the components based on density.
Next, the prepared serum is added to a microplate that is coated with antibodies specific to HBsAg. If HBsAg is present in the sample, it will bind to these antibodies.
The microplate is then incubated for a specified period, allowing time for any HBsAg present to bind to the antibodies on the plate.
After incubation, the microplate is washed multiple times. This removes any unbound substances, ensuring that only the antigen-antibody complexes remain.
A detector antibody, which is linked to an enzyme, is added next. This detector antibody binds to any HBsAg that has already attached to the plate, forming a solid complex.
Following the addition of the detector antibody, a substrate for the enzyme is applied. If HBsAg is present, the enzyme will react with the substrate, leading to a color change.
The final step is to measure the intensity of the color change using a spectrophotometer. The degree of color change corresponds to the concentration of HBsAg in the sample, allowing for quantitative assessment.
A positive result indicates the presence of HBsAg, signifying an active Hepatitis B infection, while a negative result suggests that the infection is absent. It is crucial to follow up with further tests to assess the stage and type of the infection.
The HBsAg ELISA test is a vital tool in the diagnosis of Hepatitis B. By understanding each step involved in the test, healthcare providers can ensure accurate detection and appropriate patient management.
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