Hepatitis B Surface Antigen (HBsAg) ELISA Kit

Sample

cell culture, bioprocessing solutions, and or other samples (e.g., blood, saliva, mucosa)

Species Reactivity

HBV

Detection Method

sELISA

Intended Use

The Hepatitis B Surface Antigen (HBsAg) ELISA Kit is a sandwich immunoassay for the quantification of HBsAg, recombinant or native, in cell culture, bioprocessing solutions, and or in other appropriately qualified samples from tissue fluids (e.g., blood, saliva, mucosa). The assay is not intended for the diagnosis of Hepatitis B infection.

Contents of Kit

1. Sample Diluent Concentrate (20×), 10ml
2. Wash Solution Concentrate (100×), 10ml
3. Anti-HBsAg-HRP Conjugate Concentrate (100×), 0.15ml
4. Anti-HBsAg Microwell Strip Plate, 8-well strips (12), Coated with HBsAg viral antigens, and post-coated with stabilizers.
5. HBsAg Standards, 1 ng/ml, 3 ng/ml, 6 ng/ml, 12 ng/ml, 20 ng/ml. 0.65 ml, Five (5) vials, each containing purified recombinant HBsAg with designated concentrations; diluted in buffer with protein, detergents and non-azide antimicrobials as stabilizers.
6. Positive Control [HBsAg], range on label. 0.65 ml. HBsAg of stated concentration range; diluted in buffer with protein, detergents and non-azide antimicrobials as stabilizers.
7. TMB Substrate, 12 ml. Chromogenic substrate for HRP containing TMB and peroxide.
8. Stop Solution, 12 ml. 1% sulfuric acid.

Storage

The microtiter well plate and all other reagents, if unopened, are stable at 2-8°C until the expiration date printed on the box label.

Precision

Samples containing low, medium and high concentrations of HBsAg were assayed multiple times in the same assay (n=10) to provide within-assay precision, and as duplicates in multiple assays (n=5) to obtain between-assay reproducibility.
Coefficients of variation were calculated for the concentrations using a point-to-point curve-fitting program.
HBsAg concentrations were measured with good within-assay (2.5 to 4.1 %CV) and between-assay (4.3 to 9.2 %CV) reproducibility.

General Description

Hepatitis B is an infectious disease caused by hepatitis B virus (HBV). Hepatitis, the acute illness, inflames the liver, causing jaundice, vomiting and (rarely) death. Chronic hepatitis B, however, can cause cirrhosis and liver cancer – a fatal disease. Although viral replication occurs in the liver, HBV spreads to the blood where virus-specific antigens and antibodies may be found in the infected host. Blood tests for these antigens and antibodies are used to diagnose the infection. Acute and chronic hepatitis B can be prevented by vaccination.
HBV is divided into four major serotypes (adr, adw, ayr, ayw) based on antigenic epitopes presented on its envelope proteins, and into eight genotypes (A-H) according to overall nucleotide sequence variation of the genome. Genotypes differ by at least 8% of their sequences, differences which affect severity of disease and response to treatment and possibly vaccination.
The hepatitis B surface antigen (HBsAg or Australia antigen) is the first detectable viral antigen to appear during infection, and is most frequently used to screen for the presence of infection.HBsAg is also the basis for several recent vaccines, which use synthetic recombinant HBsAgand contain no blood products.Therefore, they cannot cause HBV infection, a problem with the original vaccine prepared from plasma from patients with long-term HBV infection. Following vaccination, HBsAg may be detected in serum for several days. These vaccines have provided protection for 85-90% of individuals.Patients who developed antibodies against HBsAg (anti-HBsAg seroconversion) are usually considered non-infectious. HBsAg detection by immunoassay is used in blood screening, to establish a diagnosis of hepatitis B infection in the clinical setting (in combination with other disease markers) and to monitor antiviral treatment.

Standard Curve