ELI.H.A Amoeba

Intended Use

Amoebiasis is a parasitic infection caused by a protozoon specific to humans: Entamoeba histolytica. It’s the only pathogenic amoeba for humans. Immunological diagnosis, via the detection of antibodies, is highly important to confirm hepatic and pulmonary amoeba infections.

ELITech developed the ELI.H.A Amoeba, an indirect hemagglutination assay, for the quantitative determination of anti-Entamoeba histolytica antibodies in the serum. It is rapid, easy-to-use and easy-to-read with results available within 2 hours and reliable overnight.

Principle

Sheep red blood cells are sensitized with Entamoeba histolytica antigen.

Diluted serum is mixed with sensitized sheep red blood cells. If anti-Entamoeba histolytica antibodies are present in the serum, sensitized red blood cells will agglutinate, resulting in a cloudy red/brown deposit coating the well. In the absence of specific antibodies, sensitized red blood cells will not agglutinate, resulting in a ring-like deposit at the bottom of the well.

Non-sensitized red blood cells are also available in the kit. They ensure the specificity of the reaction by allowing identifying any interference from the natural anti-sheep agglutinins (Forssman heteroantibodies, infectious mononucleosis antibodies…).

The reaction is carried out in a U-microplate.

Each kit allows 120 tests to be carried out or 20 reactions of 6 dilutions. Results are obtained within 2 hours and are reliable overnight.

Simple Protocol

A single protocol for all our ELI.H.A kits.

1. First serum dilution
2. Microplate preparation
3. Serial dilution of serum and preparation of the ‘serum control’ well
4. Addition of sensitized RBC and non sensitized RBC
5. Gently mix the plate and let stand 2 hours before reading

Easy-to-read and easy-to-interpret results

Reagents and Material

• R1:
  Vial of 2.4 mL of sensitized red blood cells
  Quantity: 1
• R2:
  Vial of 1 mL of non-sensitized red blood cells
  Quantity: 1
• BUF:
  Vial of 55 mL of phosphate buffer pH 7.2
  Quantity: 1
• R3:
  Vial of 2 mL of adsorbent
  Quantity: 1
• CONTROL +:
  Vial of 0.2 mL of titrated positive control
  Quantity: 1
• CONTROL -:
  Vial of 0.2 mL of negative control
  Quantity: 1
• MICROPLATE:
  Microplate with a U-bottom
  Quantity: 2
• DROPPER:
  Special dropper
  Quantity: 2

Stability and Storage

• The reagents are ready-to-use.
• All the reagents are stored at 2-8°C. Do not freeze.
• The results of the stability study indicate that the product has a stability of 24 months from its manufacture.

Performance

• Diagnostic sensitivity: 93%
• Diagnostic specificity: 97%

Material required but not provided

• Automatic pipette(s) with a pipetting volume adapted to the volume that will be measured
• Contaminated waste containers
• Centrifuge
• Hemolysis tubes