Bioneovan RUO HAV-IgG ELISA is an in vitro enzyme linked immunoassay supplied for the detection of HAV-IgG in human serum or plasma. It is intended to be used as an aid in supplementary diagnosis to acute hepatitis A infection and prevalence studies among the population.

This kit uses capture ELISA method to detect anti-HAV IgG. The purified mouse anti-human IgG (μchain) antibody is coated on the solid phase of multi-wells. A conjugate HAV-Ag is added to the coated wells after diluted sample added and incubated. Then horseradish peroxidase labeled HAV-Ag is added. If HAV-IgG is present in the sample, a complex of Anti-μ-chain-HAV-IgG –HAV-Ag -labeled with HRP will form. Wash wells to remove other unbounded serum components, incubate with substrate (TMB) to form a colored product, and measure the absorbance at 450nm to indicate the presence or absence of HAV-IgG in the sample. The test is special, sensitive, reproducible and easy to operate.

Main components

1. Anti-μ-chain Coated Microwell Plate

2. Enzyme Conjugant (HAVAg-HRP)

3. Conjugate HAV-Ag

4. Negative Control Serum

5. Positive Control Serum

6. 20 X Wash Buffer (dilution prior to use)

7. Substrate A

8. Substrate B

9. Stop Solution

10. Plastic Bag

11. Seal Paper

12. Manual

TEST PROCEDURE

1. Bring ELISA Kit for Antibody IgM to Hepatitis A Virus (all reagents), and samples to room temperature before use (approximately 30 minutes).

2. Dilute concentrated wash buffer 1:19 with ddH₂O.

3. Dilute the sample (1:1000) with physiological saline.

4. For each test, set one blank, two positive and three negative controls. Add 100μl positive and negative control serum into positive and negative control wells respectively.

5. Add 100μl diluted sample into other test wells.

6. Cover wells with seal paper, then incubate 30 minutes at 37°C.

7. Discard the liquid in all wells and fill the wells with wash solution. Lay aside for 15 seconds, discard the liquid in all wells and fill the wells with wash solution. Repeat 5 times and dry wells after last wash.

8. Add 50 μl conjugate HAV-Ag in each well except the blank.

9. Addμl 50 μl Enzyme Conjugant in each well except the blank

10.Cover wells with seal paper, then incubate 30 minutes at 37°C.

11. Repeat step 7.

12.Add 50 μl substrate A and B respectively to each well,mix gently protected from light and incubate 15 minutes at 37°C.

13. Add 50μl of stop solution into each well to stop the reaction, including blank well.

14. Measure the absorbance at 450 nm against the blank, or measure the absorbance at 450 nm/630-690 nm.