For in vitro diagnostic and professional use only.
Name
Anti-Ct IgG ELISA Test Kit (Serum/Plasma)
INTENDED USE
This kit is a qualitative detection of human serum/plasma of Chlamydia trachomatis (Ct) IgG. The kit is
suitable for clinical screening and diagnosis of Ct infection in serum/plasma.
TEST PRINCIPLE
Ct antigen is absorbed in solid phase to the polystyrene reaction microplate. If there is Ct IgG antibody
in test sample, it binds to Ct antigen coated in microplate, forms antigen-antibody complex, and then
binds to the enzyme labeled anti-antibody and forms antigen-antibody-antibody complex on surface
of the microplate, and display blue color in corresponding well via the action of substrate. Therefore, it
can detect specifically the Ct IgG in human serum/plasma.
TEST PROCEDURE
1. Equilibrate all reagents to room temperature for 15 minutes prior to use.
2. Dilute the wash buffer at the rate of 1:40 dilution with distilled water before use.
3. Add 100μL Sample Diluent in the corresponding well (Do not add in the blank well, negative
control wells and positive control well.) The sample should be corresponding to the number of micro
plate, each plate should be provided with negative control 2 wells, positive control 1 well and blank
control 1 well. Add 5μL sample in the corresponding well, mix thoroughly by using the pipette, add
50μL negative control and positive control to negative control wells and positive control well (Do not
add in the blank well).
Note: Use a separate disposal pipette tip for each specimen, Negative and Positive Control to
avoid cross contamination.
4. Shake gently to mix for 30s. Incubate at 37°C for 20 minutes with the sealing plate membrane
sealing the plate.
5. At the end of the incubation, remove and discard the plate cover. Take out, add wash buffer to
each well for 20 seconds. Repeat 5 times. After the final washing cycle, turn the plate over onto
blotting paper or clean towel, and tap it to remove any remainders.
6. Respectively adding Enzyme Conjugate 50µL (Do not add in the blank well)
7. Incubate at 37°C for 20 minutes with the sealing plate membrane sealing the plate. Repeat the
wash step for 5 times as in step 5.
8. Add Substrate A 50µL and Substrate B 50µL (Do not add in the blank well). Incubate at 37 ℃ for
10 minutes with the sealing plate membrane sealing the plate.
9. Add 50μL Stop Solution to each well (Do not add in the blank well). Mix gently by shaking, read the
absorbance within 10 minutes after stopping the reaction. Calibrate the plate reader with the Blank
well and read the absorbance at 450nm.If a dual filter instrument is used, set the reference
wavelength at 630nm. Set no blank well is allowed if use dual wavelength to detect. Calculate the
Cut-off value and evaluate the results.
INTERPRETATION OF RESULTS
Chronometry: Read the sample's optical density (OD) at 450nm with a micro plate reader.
The Mean negative control OD value ≤ 0.1 and Mean positive control OD value ≥ 0.8, the test is valid,
otherwise the test is invalid.
Cut-Off value (C.O.) = The Mean OD value of negative control x 3 (Calculated by 0.10 when the Mean
negative control OD value is < 0.10, calculated by actual value when the Mean negative control OD
value is > 0.10)
Positive Results: Sample OD value ≥ C.O.
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