HBsAg ELISA kit, which is an enzyme-linked immunosorbent assay for the qualitative detection of HBsAg in human serum or plasma. The HBsAg ELISA Kit is designed as an aid in screening blood donors, and aid in the diagnosis of clinical conditions related to hepatitis B-infected individuals.
Materials & Instrumentations Required not provided:
1. Freshly distilled or deionized water 2. Dispensing system and/or pipette & tips 3. EIA kit Microplate Washer 4. EIA kit Microplate Reader, single wavelength 450nm or dual-wavelength 450nm & 630nm
HBsAg ELISA Assay Kit Principle
The HBsAg ELISA Test kit employs an antibody sandwich ELISA technique where monoclonal antibodies unique to HBsAg, are pre-coated on polystyrene microwell strips. The serum or plasma sample is added together with a second antibody, the HRP Conjugate, (horseradish peroxidase), and directed against a different epitope of HBsAg. Throughout the time of incubation, a specific immunocomplex that may have formed (indicating the presence of HBsAg) is captured in the solid phase. After washing, to eliminate serum proteins and unbound HRP-conjugate, chromogen solutions containing tetramethyl-benzidine (TMB) and urea peroxide are added to the wells. Next, the colorless chromogens are hydrolyzed by the bound HRP-conjugate to a blue-colored product while in the presence of the antibody-antigen-antibody (HRP) sandwich immunocomplex. Halting the reaction with sulfuric acid, the blue color then turns yellow. The color intensity can be gauged proportionally to the amount of antigen captured in the wells, and to the amount in the sample, respectively. The wells remain colorless if the HBsAg result is negative. At SOUTH PUNJAB LAB this test is done by VIDAS.
HBsAg ELISA Test Results Interpretations
(S = the individual optical density (OD) of each specimen) Negative Results (S/C.O. <1): samples giving an optical density less than the Cut-off values are considered negative, which indicates that no hepatitis B virus surface antigen has been detected with this HBsAg ELISA, therefore the patient is probably not infected with the hepatitis B virus. Positive Results (S/C.O.>1): samples giving an optical density greater than or equal to the Cut-off value are considered initially reactive, which indicates that HBV surfaces antigen has probably been detected with this HBsAg ELISA. For additional details please refer to the instructions for use.
HBsAg ELISA Assay Performance Characteristics
Analytical Specificity 1. No cross-reactivity observed with samples from patients infected with HAV, HCV, HIV, CMV, and TP 2. No interference from rheumatoid factors up to 2000U/ml was observed 3. No high dose hook effect up to HBsAg concentrations of 200000ng/ml observed during clinical testing Reference: http://www.rapidtest.com/
