Hepatitis A Virus elisa test kit is an in vitro enzyme immunoassay for the detection of HAV-IgM in human serum or plasma.

Hepatitis A Virus elisa test  kit uses capture ELISA method to detect anti-HAV IgM. The purified mouse anti-human IgM (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 peroxides labeled HAV-Ag is added. If HAV-IgM is present in the sample, a complex of Anti-μ-chain-HAV-IgM –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-IgM in the sample. The test is special, sensitive, reproducible and easy to operate.

Blood serum samples are routinely prepared form vein. Blood plasma samples are routinely prepared with routine amount of anticoagulant such as heparin or sodium citrate. Sample can be stored at 4°C if tested within five days. Sample can be stored at -20°C at least for 3 months. Avoid hemolysis and repetitive freeze and thaw of samples. Samples with cloud or precipitation should be centrifugated or filtered before test. Prevent serum from bacterium contamination during collecting and storing.

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, and 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 50μl conjugate HAV-Ag in each well except the blank.

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

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

11. Repeat step 7.

12. Add 50μl 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.

The testing is for qualitative and assistant diagnosis. Confirmation of infection should refer to the clinical and other diagnosis.

If the OD of positive controls is not below 1.0, OD of negative is not higher than 0.1, the assay result is validated. Otherwise, repeat the test.

1. 1.       The samples should be fresh, avoid hemolysis, bacteria growing and repetitive freeze and thaw.
2. 2.       Do not interchange reagents between kit lots. The seal paper can’t be used repeatedly.
3. 3.       Mix reagents well before use. If crystal form in certain reagents, such as wash buffer, it can be used after warming up and mixing well.
4. 4.       Follow instruction exactly during assay, especially in temperature and time for reactions. All pipetting devices should be used with care and calibrated regularly following the manufacturer’s instructions.
5. 5.       Put the remained reagents to the sealed pouch, and return to 2~8°C in time.
6. 6.       To prevent cross-contamination, wear gloves and working suits throughout the procedure, and execute the disinfection and isolation regulations strictly. Dispose of all samples and materials used to perform the test. The 5.0g/L liquid sodium hypochlorite solution or 121°C high pressure steam may be used to disinfect samples and materials before disposal (The positive control serum in the kit has been inactivated already)