Specifications

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HBeAb ELISA Kit

CatalogNo.:BE104A

 

Enzyme-linked immunosorbent assay for the detection antibody against HBeAg in serum or plasma

INTENDED USE

 

Anti-HBeAg antibody (HBeAb) EIA is a qualitative enzyme immunoassay for the detection of antibody to e antigen of hepatitis B virus (HBe) in human serum or plasma.

 

SUMMARY AND PRINCIPLE OF THE TEST

 

The presence of antibody against hepatitis B viral e antigen is used as an indicator for (1) early HBs antigenemia before the peak of viral replication and (2) early convalescence when HBeAg has declined below detectable levels. It is also useful to confirm a seroconversion. The seroconversion from HBeAg positivity to anti-HBe positivity indicates a reduced level of infectious virus because virus replication has decreased.

 

Anti-HBe antibody test is a competitive enzyme immunoassay in which anti-HBe antibodies from specimens compete with a constant amount of Horseradish Peroxidase (HRP) conjugated anti-HBe antibody for a limited number of HBeAg in the neutralizing reagent added to the well. The microtiter well is coated with polyclonal antibodies against HBeAg as catcher for HBeAg. A serum specimen is added to the microtiter wells together with HRP conjugated anti-HBe-HBeAg complex. After incubation, anti-HBe antibodies in specimen, if present, compete with constant amount of HRP- conjugated anti-HBe for limited amount of HBeAg added into the wells. The unbound enzyme conjugates will be washed away and the chromogen substrate solution containing hydrogen peroxide is added to the wells for color development. Thus, the amount of HRP-conjugated anti-HBe bound to the well is inversely proportional to the concentration of anti-HBe antibody in the specimen. The absorbance of controls and specimens is determined using EIA reader with wavelength set at 450 nm (450 nm/630 nm).

 

REAGENTS

 

Materials provided with the kits:

1. Microtiter Well coated with purified anti-HBe: 96 tests
2. Negative Control: One vial of 0. 5ml anti-HBe Negative Control.
3. Positive Control: One vial of 0. 5ml anti-HBe Positive Control.
4. Enzyme Conjugate: 6 ml containing HRP-conjugated-anti-HBe-HBeAg complex for 96 tests.
5. Wash Buffer Concentrate (20 x): 20 ml for 96 tests. The buffer should be diluted 20 times with distilled water before use.
6. Substrate Solution A: 6 ml HRP Substrate for 96 tests.
7. Substrate Solution B: 6 ml TMB Chromagen Substrate for 96 tests.
8. 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.

PRECAUTION FOR USERS

1. For in-vitro diagnostic use only.
2. Must not use kit beyond the expiration date.
3. Do not mix components from kits with different lot number.
4. Avoid microbial contamination of reagents.
5. Do not pipet reagent by mouth and no smoking or eating while performing assays.
6. Wear gloves during the whole process and avoid reagents or specimen spilling-out.
7. Wipe up the spills using 5% hypochlorite solution.
8. Decontaminate all liquids or solid wastes before deposing.

 

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