HBsAg ELISA Kit (96 Tests)

HBsAg ELISA Kit (96 Tests) is an in vitro enzyme immunoassay for the qualitative detection of Hepatitis B surface antigen (HBsAg) in human serum or plasma. It is based on a direct, non-competitive, solid-phase enzyme immunoassay using horseradish peroxidase (HRP) as the marker enzyme. For broader background on hepatitis B and HBsAg testing, see the WHO hepatitis B fact sheet and CDC guidance on clinical testing and diagnosis for hepatitis B. As part of a wider ELISA product range, this kit can also be explored alongside the HCV ELISA Kit and HIV ELISA 4th Gen Kit.

Pack size: 96 Tests
Storage: 2°C to 8°C
Expiry: 18 months

Product Overview

The HBsAg ELISA Kit is designed for the qualitative detection of HBsAg in human serum or plasma. During the assay, HBsAg in the sample binds to anti-HBs antibodies coated on the microwell. An HRP-conjugated anti-HBs antibody then binds to the complex. After washing, TMB substrate develops colour, the reaction is stopped, and absorbance is measured at 450 nm with 630 nm as the reference wavelength.

Product Specifications

• Test type: Microwell ELISA
• Application: Qualitative detection of HBsAg
• Sample type: Human serum or plasma
• Principle: Direct, non-competitive, solid-phase enzyme immunoassay
• Marker enzyme: Horseradish peroxidase (HRP)
• Read wavelength: 450 nm
• Reference wavelength: 630 nm
• Pack size: 96 Tests
• Storage: 2°C to 8°C
• Sensitivity / detection limit: Analytical sensitivity 0.4 PEIU/ml
• Diagnostic sensitivity: 100%
• Diagnostic specificity: 100%

Intended Use

HBsAg Microwell ELISA is an in vitro enzyme immunoassay for the qualitative detection of Hepatitis B surface antigen (HBsAg) in human serum or plasma.

Principle of the Test

HBsAg Microwell ELISA is a direct, non-competitive, solid-phase enzyme immunoassay with HRP as the marker enzyme.

1. If present, HBsAg in the sample binds to anti-HBs antibodies coated on the microwell.
2. An HRP-conjugated anti-HBs antibody binds to the complex.
3. After washing, TMB substrate produces a colour change.
4. The reaction is stopped and absorbance is measured.

Kit Contents

1. HBsAg antibody coated microstrips (1 × 96 wells)
2. Sample diluent (1 × 6.0 ml)
3. Negative control (1 × 0.5 ml)
4. Positive control (1 × 0.5 ml)
5. Conjugate concentrate (101x) (1 × 0.3 ml)
6. Conjugate diluent (1 × 15 ml)
7. TMB substrate concentrate (101x) (1 × 0.3 ml)
8. Substrate buffer (1 × 15 ml)
9. Wash solution concentrate (20x) (1 × 30 ml)
10. Stop solution (1 × 6 ml)
11. Microstrip covers (02 nos.)
12. Product insert (01 no.)
13. Procedure card (01 no.)

Specimen Requirements and Storage

• Specimen: Human serum or plasma
• Store specimens at 2–8°C.
• If specimens are not tested within 7 days, store frozen at -15°C or colder.
• Avoid multiple freeze–thaw cycles.
• After thawing, mix specimens thoroughly before testing.

How to Use the Kit

1. Bring the required reagents and microwells to room temperature before use.
2. Add sample and controls as described in the kit procedure.
3. Incubate according to the IFU method.
4. Wash the wells as instructed.
5. Add conjugate and continue the procedure as per the IFU.
6. Add TMB substrate and allow colour to develop.
7. Stop the reaction and read absorbance at 450 nm with 630 nm as reference wavelength.
8. Interpret results using the calculated cut-off value.

Interpretation of Results

Cut-off value (COV)

1. Calculate the negative control mean (NCx)
2. COV = NCx + 0.100

Result interpretation

• Reactive: Sample absorbance ≥ COV
• Non-reactive: Sample absorbance < COV
• Grey zone: Absorbance within ±10% of COV

Retesting note

If samples are considered reactive, the test should be conducted two more times.

Quality Control

Results are valid if the following control criteria are met:

• Blank absorbance < 0.050
• Mean absorbance of negative controls < 0.100
• Mean absorbance of positive controls > 1.000

If results fall outside these ranges, the test should be conducted again.

Performance Characteristics

• Analytical sensitivity / detection limit: 0.4 PEIU/ml
• Diagnostic sensitivity: 100%
• Diagnostic specificity: 100%

Limitations

• Handle all specimens as potentially infectious.
• Do not use reagents beyond the stated expiry date.
• Do not modify the test procedure or substitute reagents unless stated as interchangeable.
• Do not allow wells to become dry during the assay procedure.
• Samples in the grey zone require careful retesting as described in the IFU.

FAQs

What is the HBsAg ELISA Kit used for?

It is used for the qualitative detection of Hepatitis B surface antigen (HBsAg) in human serum or plasma. For general background on hepatitis B and how HBsAg testing fits into diagnostic workflows, refer to the WHO hepatitis B fact sheet and CDC hepatitis B testing guidance.

What sample type is required for the HBsAg ELISA Kit?

The required sample type is human serum or plasma.

How is the HBsAg ELISA Kit read?

Absorbance is measured at 450 nm with 630 nm as the reference wavelength.

What is the sensitivity or detection limit of the HBsAg ELISA Kit?

The analytical sensitivity stated in the IFU is 0.4 PEIU/ml.

How are results interpreted in the HBsAg ELISA Kit?

Results are interpreted against a calculated cut-off value (COV). Samples may be reactive, non-reactive, or in the grey zone depending on absorbance relative to the cut-off.

What should be done if a sample is reactive?

If a sample is considered reactive, the test should be conducted two more times.