Toxocara Canis ELISA Kit

Catalog No. ABIN997049

Product Details

Target: Toxocara Canis

Reactivity: Dog Roundworm (Toxocara canis)

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Application: ELISA

Purpose: For the qualitative screening of IgG antibodies to Toxocara in serum and plasma using an Enzyme-Linked Immunosorbent Assay (ELISA) technique.

Sample Type: Serum

Analytical Method: Qualitative

Specificity: 100 %

Material not included: 1. Pipettes
2. Squeeze bottle for washing strips (narrow tip is recommended)
3. Reagent grade water and graduated cylinder
4. Tubes for sample dilution
5. Absorbent paper
6. 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
Limitations of procedure: Serologic results are an aid in diagnosis but cannot be used as the sole method of diagnosis. ISO 13485 ISO 9001 Diagnostic Automation/Cortez Diagnostics, Inc. 23961 Craftsman Road, Suite E/F, Calabasas, California 91302 USA Date Adopted

Sample Volume: 5 μL

Assay Time: 1 h

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 the serum from the clot. Serum should be stored at 2 - 8 °C if it is to be analyzed within a few days. Serum may be held for 3-6 months by storage at -20 °C or lower. 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 mL sera and 315 mL dilution buffer). chromogen (tetramethylbenzidine or TMB) is added. With the presence of Enzyme Conjugate and the peroxidase causing the consumption of peroxide, the chromogen changes to a blue color. The blue color turns to a bright yellow color after the addition of the stop solution, which ends the reaction. ELISA readers can be used to obtain results, or the reaction can be ready visually. Test Toxocara IgG ELISA Method Enzyme Linked Immunosorbent Assay Principle Sandwich Complex Detection Range Qualitative Positive, Negative Control Sample 5 μL serum Total Time - 20 min. Shelf Life 12 Months from the manufacturing date Specificity 100 %

Assay Procedure:

1. Break off number of wells needed (two for controls plus number of samples) and place in strip holder.
   2. Add 100 mL (or two drops) of the negative control to well #1, 100 mL of the positive control to well #2 and 100 mL 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.
   5. Add100 mL 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. Slap plates against paper toweling to remove excess moisture.
   8. Add 100 μL of the Chromogen to every well. cc
   9. Incubate at room temperature for 5 minutes.
   10. 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 the Test 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.\
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

Storage: 4 °C

Expiry Date: 12 months

Target Details

Target: Toxocara Canis

Target Type: Species

Background: Toxocariasis is the infection caused by roundworm of the genus Toxocara (usually T. canis, rarely T. cati) and is acquired by ingesting soil contaminated with embryonated eggs from an animal’s feces. These eggs become embryonated after 2 to 5 weeks after being passed by the animal. Thus, human infection does not occur by contact with fresh feces. In addition, infected humans cannot pass the infection to other humans. 6 The disease manifests itself as visceral larval migrans (VLM) and ocular larval migrans (OLM). Signs and symptoms of VLM may vary from an asymptomatic state with mild eosinophilia to a severe and potentially fatal disorder. Patients with OLM also vary widely in presentation, from acute lesions in the eye to asymptomatic infections. Toxocara larva migrans is believed to be the second most common helminth infection in developed countries.