HBeAg ELISA test detect Hepatitis B E antigen,96wells/kit, Elisa Sandwich method, for quantitative measurement

1. HBeAg EIA is a qualitative enzyme immunoassay for the detection of HBeAg
2. human serum or plasma.
3. in vitro diagnostic kit

SUMMARY

This kit uses “ sandwich” ELISA method in which, polystyrene microwell strips are pre-coated with monoclonal antibodies specific to HBeAg. Patient’s serum or plasma sample is added to the microwell together with a second monoclonal antibody conjugated to horseradish peroxidase (HRP-Conjugate).During incubation, the specific immunocomplex formed in case of presence of HBeAg in the sample is captured on the solid phase. After washing to remove sample and unbound HRP-Conjugate, Chromogen solutions containing Tetramethylbenzidine (TMB) and urea peroxide are added into the wells. In presence of the antibody-antigen-antibody(HRP) “sandwich “ complex, the colorless Chromogens are hydrolyzed by the bound HRP conjugate to a blue-colored product. The blue color turns yellow after stopping the reaction with sulfuric acid. The amount of color intensity can be measured and is proportional to the amount of antigen captured in the wells, and to the sample respectively. Wells containing samples negative for HBeAg remain colorless.

PRINCIPLE

This kit is an enzyme -linked immunosorbent assay (ELISA) for qualitative detection of hepatitis B virus e antigen (HBeAg) in human serum or plasma. It is intended for use in clinical laboratories for diagnosis and management of patients related to infection with hepatitis B virus. HEPATITIS B VIRUS E ANTIGEN ELISA. One-Step Incubation, Double Antibody Sandwich Principle

REAGENT

Materials provided with the kits:

1. Negative Control: 0.5ml HBeAg Negative Control.
2. Positive Control: 0.5ml HBeAg Positive Control.
3. Enzyme Conjugate:.6 ml containing HRP-conjugated-anti-HBe
4. Wash Buffer Concentrate (20 x): 20 ml for 96 tests. The buffer should be diluted 20 times with distilled water before use.
5. Substrate Solution A: 6 ml HRP Substrate for 96 tests.
6. Substrate Solution B: 6 ml TMB Chromagen Substrate for 96 tests.
7. Stop Solution: One bottle of 6 ml 2N Sulfuric Acid

Materials required but not provided:

1. Precision pipettes: 0.02, 0.05, 0.10, 0.15, 0.20, and 1.0 ml.
2. Disposable pipette tips.
3. Distilled water.
4. Humidified Box capable of maintaining 37°C
5. Absorbent paper or paper towel.
6. Micro titer plate or strip-well washer
7. Micro titer plate reader.