HEV IgM Elisa Test Kit Serum Vs Plasma For Antibody Detection

Diagnositc Kit for IgM Antibody to Hepatitis E Virus(ELISA)
Catalog No.:BE402A

1. PRINCIPLE

This kit employs solid phase, capture ELISA method for detection of IgM antibodies to HEV (anti-HEV) in serum or plasma with two-step incubation procedure. Polystyrene microwell is pre-coated with purified activated mouse anti human IgM (μ chain) monoclonal antibody. The HRP conjugated recombinant HEV antigen serves as tracer. TMB is substrate for HRP. The enzyme reaction with substrate TMB produces a color change, and the intensity of the absorbance at 450 nm indicates the presence or absence of Anti-HEV antibodies IgM in the sample. The test is specific, sensitive, reproducible and easy to operate. It is for diagnosis of early infection and epidemic survey.

2. MATERIALS PROVIDED

1. Antigen Coated Micro well Plate 1 block (96wells)
2. Specimen Diluents 1 bottle (12ml)
3. Enzyme Conjugant (anti human IgM -HRP) 1 bottle (12ml)
4. Negative Control 1 vial (1ml)
5. Positive Control 1 vial (1ml)
6.20 X Wash Buffer (dilution prior to use) 1 bottle (30ml)
7. Substrate A 1 bottle (6ml)
8. Substrate B 1 bottle (6ml)
9. Stop Solution (2M H2SO4) 1 bottle (6ml)
10. Plastic Bag 1 bag
11. Seal Paper 3 pieces
12. Manual 1 each

3TEST PROCEDURE

1. Bring ELISA Kit for Antibody IgG to Hepatitis E Virus (all reagents), and Specimens to room temperature before use (approximately 30 minutes).

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

3. 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.

4. Add 100μl sample diluents in each other test well, then at 10 μl test specimen into the test wells. Pipet up and down to mix the samples well.

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

6. 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.

7. Add 100 μl Enzyme Conjugant in each well except the blank.

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

9. Repeat step 6.

10. Add 50μl substrate A and B respectively to each well including the blank well. Mix gently, protected from light and incubates 15 minutes at 37°C.

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

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