Entamoeba Ag ELISA Kit

Sample

Stool

Species Reactivity

Human

Detection Method

sELISA

Intended Use

The Entamoeba ELISA is an enzyme-linked immunosorbent assay (ELISA) for the qualitative identification of Entamoeba histolytica and Entamoeba dispar in human stool samples. For Research Use Only. Not for use in diagnostic procedures.

Contents of Kit

1. Plate 1: 96-well microwell plate, 12 microwell strips (which can be divided) in strip holder, coated with specific monoclonal antibodies (mouse) to Entamoeba histolytica and Entamoeba dispar
2. Diluent 1: 100 ml Sample dilution buffer, protein-buffered NaCl solution; ready for use; blue colored
3. Wash Buffer: 100 ml Wash buffer, phosphate-buffered NaCI solution (concentrated 10 fold); contains 0.1 % thimerosal
4. Positive Control: 2 ml Inactivated Entamoeba antigen; ready for use
5. Negative Control: 2 ml Negative control (sample dilution buffer); ready for use
6. Conjugate 1: 13 ml Biotin conjugated antibodies (mouse) to Entamoeba antigens in stabilized protein solution; ready for use, green colored
7. Conjugate 2: 13 ml Streptavidin poly-peroxidase conjugate in stabilized protein solution; ready for use; orange colored
8. Substrate: 13 ml Hydrogen peroxide/TMB; ready for use
9. Stop: 12 ml Stop reagent; 1 N sulfuric acid; ready for use
10. Plate Sealer 2

Storage

All reagents must be stored at 2 – 8 °C and can be used until the date printed on the label.
Providing the 1× working wash buffer is stored at 2 – 8 °C, it can be used for a maximum of 4 weeks. Microbial contamination must be prevented. After the expiry date, the quality guarantee is no longer valid.
The aluminum bag must be opened with scissors in such a way that the clip seal is not torn off.
Any microwell strips which are not required must be returned to the aluminum bag and immediately stored at 2 – 8 °C.
The colorless substrate must be protected from direct light to prevent it from decomposing or turning blue due to auto-oxidation. Once the substrate has turned blue, it must not be used.

Precision

To determine intra-assay reproducibility, 40 replicates of these references were assayed, representing the complete measurement range from negative to high-positive. The mean values and the coefficients of variation (CV) were determined for three lots of the kits. For the interassay reproducibility, references from 10 different working days were assayed in duplicates, with two runs per day. The measurements were determined by three technicians for three lots of the kits. The inter-lot reproducibility was determined for all three lots of the kits.

Sensitivity

To determine the analytic sensitivity of the Entamoeba ELISA, a linear dilution series from a sample with a known quantity of Entamoeba cysts was produced and then measured in triplicates. The limit of detection (LoD) is the last concentration to be evaluated as positive in all replicates.