Cysticercosis ELISA 试剂盒

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: Positive - 0.5 OD units and above. Negative - Absorbance reading 0-0.3 OD units.
Limitations of procedure: Serologic results are an aid in diagnosis but cannot be used as the sole method of diagnosis. Significant cross reactions with Echinococcus infections will occur in this assay. If Echinococcus infection cannot be ruled out in the differential diagnosis, a positive sample should be confirmed by other means (i.e. immunoblot offered by the CDC) or by other non-serological means.

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.

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 to 6 months by storage at -20 °C or lower. Lipemic and strongly hemolytic serum should be avoided. Do not heat inactive serum and avoid repeated freezing and thawing of samples. Freeze sample at -20 °C or lower if not used immediately. 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). 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 results can be read visually. Test Cysticercosis IgG (T. solium)ELISA Method Enzyme Linked Immunosorbent Assay Principle Sandwich Complex Detection Range Semi-quantitative : Positive, Negative Sample 5 μL Total Time - 20 min. Shelf Life 12 Months from the manufacturing date Specificity 100 %

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.
   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. Slap wells against cc paper towels to remove excess moisture.
   8. Add 100 μL of the Chromogen to every well.
   9. Incubate at room temperature for 5 minutes.
   10. Add 100 μL of the Stop Solution and mix by tapping strip holder.

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.
Reference Method 1
- 0 65 Positive Agreement: 100 % (15/15) Negative Agreement: 98.4 % (65/66)
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.