Echinococcus ELISA Kit

Catalog No. ABIN997044

Product Details

Target: Echinococcus

Reactivity: Echinococcus granulosus

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Application: ELISA

Purpose: The Echinococcus ELISA Test is For the qualitative screening of serum IgG antibodies to Echinococcus sp. using an Enzyme-Linked Immunosorbent Assay (ELISA) technique

Analytical Method: Qualitative

Specificity: 100 %

Material not included: 1. Pippettes
2. Squeeze bottle for washing strips (narrow tip is recommended)
3. Reagent grade water and graduated cylinder
4. Tubes for sample dilution
5. Sample Dilution Tubes
6. Absorbent paper
7. ELISA plate reader with a 450 nm and a 620-650 nm filter (optional if results are read visually).

Application Details

Comment:

Quality Control:
The use of controls allows validation of kit stability. The kit should not be used if any of the controls are out of range. Expected values for the controls are: Negative - 0.0 to 0.3 OD units Positive - 0.5 OD units and above
1. Do not use solutions if they precipitate or become cloudy. Wash concentrate may show crystallization upon storage at 2-8 °C. Crystallization will disappear after dilution to working strength.
2. Do not use serum that may have supported microbial growth, or is cloudy due to high lipid content. Samples high in lipids should be clarified before use.
3. Treat all sera as if capable of being infectious. Negative control has been tested and found negative for Hepatitis B surface antigen and for the antibody to HIV by required test methods. This product should be used under appropriate safety conditions that would be used for any potentially infectious agent.
4. Do not add azides to the samples or any of the reagents.
Limitations of procedure: Serologic results are an aid in diagnosis but cannot be used as the sole method of diagnosis.

Plate: Pre-coated

Reagent Preparation:

Preparation Wash Buffer - Remove cap and add contents of bottle to 475 mL of reagent grade water. Place diluted wash buffer into a squeeze bottle with a narrow tip opening.
Note:
Washings consist of filling to the top of each well, shaking out the contents and refilling. Avoid generating bubbles in the wells during the washing steps.

Sample Preparation:

Serological specimens should be collected under aseptic conditions. Coagulate blood and remove serum. Hemolysis is avoided through prompt separation of serum from the clot. Serum should be stored at 2-8 °C if it is to be used within a few days. Serum may be held for 3 to 6 months by storage at -20° C or lower. Lipemic and strongly hemolytic serum should be avoided. Do not heat inactivate serum and avoid repeated freezing and thawing of samples. Test samples: Make a 1:64 dilution of patients' sera using the dilution buffer (e.g. 5 μL sera and 315 μL dilution buffer).

Assay Procedure:

1. Break off number of wells needed (two for controls plus number of samples) and place in strip holder.
   2. Add 100 μL (or two drops) of the negative control to well #1, 100 μL of the positive control to well #2 and 100 μL of the diluted (1:64) test samples to the remaining wells. Note: Negative and positive controls are supplied prediluted. Do not dilute further.
   3. Incubate at room temperature for 10 minutes.
   4. Shake out contents and wash 3 times with the diluted wash buffer. cc
   5. Add 100 μL of Enzyme Conjugate to each well.
   6. Incubate at room temperature for 5 minutes.
   7. Shake out contents and wash 3 times with wash buffer.
   8. Slap wells vigorously against paper towels to remove excess moisture
   9. Add 100 μL of the Chromogen to every well.
   10. Incubate at room temperature for 5 minutes.
   11. Add 100 μL of the Stop Solution and mix by tapping strip holder.

Calculation of Results:

Visually: Look at each well against a white background (e.g. paper towel) and record as clear or +, ++ or +++ reaction. ELISA Reader: Zero reader on air. Set for bichromatic readings at 450/620-650 nm. Troubleshooting Negative control has excessive color after development. Reason: inadequate washings Correction: wash more vigorously. Remove excessive liquid from the wells by tapping against an Absorbent towel. Do not allow test wells to dry out. Interpretation of Results - ELISA Reader Zero ELISA reader on air. Read all wells at 450/650-620 nm. Positive - Absorbance reading equal to or greater than 0.3 OD units. Negative - Absorbance reading less than 0.3 OD units. A negative OD reading indicates that the patient has no detectable level of antibodies. This may be due to lack of infection or poor immune response by the patient. Interpretation of Results -Visual Compare results to the controls. A sample should be interpreted as positive if the degree of color development is significant and obvious.
Negative Agreement: 100 % (65/65) *Reference Method refers to a commercially available ELISA.
The number of individuals showing positive results can vary significantly between populations and geographic regions. If possible, each laboratory should establish an expected range for its patient population.

Restrictions: For Research Use only

Handling

Target Details

Target: Echinococcus

Target Type: Bacteria