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TriiodothyronineT3 ELISA test , high accuracy, Elisa Sandwich method, for quantitative measurement
Product Name: TriiodothyronineT3 ELISA test
Intended Use:
This Total T3 EIA is intended for the quantitative determination of triiodothyronine (T3) concentration in human serum. This test is useful in the diagnosis and treatment of thyroid diseases such as hyperthyroidism.
Summary:
Measurement of serum triiodthyronine concentration is generally regarded as a valuable tool in the diagnosis of thyroid dysfunction. This importance has provided the impetus for the significant improvement in assay methodology that has occurred in the last two decades. The advent of monospecific antiserum and the discovery of blocking agents to the T3 binding serum proteins has enabled the development of procedurally simple radioimmunoassays
Principle of Test:
The Total Triiodthyronine (T3) ELISA Kit is a solid phase enzyme-linked immunosorbent assay (ELISA) based on the principle of competitive binding. The microtiter wells are coated with a polyclonal goat-anti-mouse antibody. Standards, controls and patient serum incubate together with Assay Reagent containing monoclonal anti-T3 antibodies. In the following incubation with Conjugate the endogenous T3 of a patient sample competes with the T3-horseradish peroxidase conjugate for a limited number of insolubilized binding sites. After incubation the unbound conjugate is washed off. The amount of bound peroxidase conjugate is inversely proportional to the concentration of T3 in the sample. After addition of the substrate solution, the intensity of colour developed is inversely proportional to the concentration of T3 in the patient sample.
Kit Components
* contains 0.03 % ProClin 300
** contains 0.03 % ProClin 300 + 0.01 % Gentamicin sulphate
*** contains 0.03 % ProClin 300 + 0.015 % 5-bromo-5-nitro-1,3-dioxane (BND) + 0.010 % 2-methyl-2H-isothiazol-3-one (MIT)
ASSAY PROCEDURE
1. Secure the desired number of Microtiter wells in the frame holder.
2. Dispense 10 µL of each Standard, Control and samples with new disposable tips into appropriate wells.
3. Incubate for 5 minutes at room temperature (18 °C – 25 °C).
4. Dispense 100 µL Enzyme Conjugate into each well.
Thoroughly mix for 10 seconds. It is important to have a complete mixing in this step.
5. Incubate for 80 minutes at room temperature (18 °C – 25 °C).
6. Briskly shake out the contents of the wells.
Rinse the wells 5 times with diluted Wash Solution (400 µL per well). Strike the wells sharply on absorbent paper to remove residual droplets.
PERFORMANCE CHARACTERISTICS
1. Sensitivity
The sensitivity of this Total T3 EIA test is defined as the lowest concentration of T3 that can be distinguished from 0 ng/mL as calculated from the 95% confidence limits of the 0 ng/mL standard absorbance. This method will reliably detect T3 concentrations as low as 0.2 ng/ml.
2. Precision
a. Intra-Assay Precision
Within-run precision was determined by replicate determinations of three different control sera in 1 assay.
| Serum Sample | 1 | 2 | 3 |
| Number of Replicates | 20 | 20 | 20 |
| Mean T3 (ng/mL) | 0.85 | 2.48 | 4.41 |
| Standard Deviation | 0.08 | 0.10 | 0.14 |
| Coefficient of Variation (%) | 9.6% | 4.1% | 3.2% |
b. Inter-Assay Precision
Between-run precision was determined by replicate measurements of three different serum samples over a series of individually calibrated assays.
| Serum Sample | 1 | 2 | 3 |
| Number of Replicates | 20 | 20 | 20 |
| Mean T3 (ng/mL) | 0.79 | 2.53 | 4.10 |
| Standard Deviation | 0.08 | 0.08 | 0.06 |
| Coefficient of Variation (%) | 10.3% | 3.2% | 1.4% |
3. Recovery and Linearity Studies
a. Recovery
Various patient serum samples of known T3 levels were combined and assayed in duplicate. The mean recovery was 106%.
| Expected Conc. (ng/mL) | Observed Conc. (ng/mL) | % Recovery |
| 5.05 | 4.1 | 99% |
| 2.71 | 2.73 | 101% |
| 1.46 | 1.52 | 104% |
| 0.68 | 0.65 | 96% |
| Mean Recovery #1 = 100 % | ||
| 5.81 | 6.16 | 106% |
| 3.02 | 3.17 | 105% |
| 1.54 | 1.75 | 114% |
| 0.71 | 0.84 | 119% |
Mean Recovery #2 = 111 %
Two samples were serially diluted with zero T3 standard to determine linearity. The mean recovery was 99%.
| # | Dilution | Expected Conc. (ng/mL) | Observed Conc. (ng/mL) | % Expected | ||
| 1. | Undiluted | ---- | 6.06 | ---- | ||
| 1:2 | 3.03 | 3.05 | 101% | |||
| 1:4 | 1.51 | 1.46 | 97% | |||
| 1:8 | 0.75 | 0.68 | 91% | |||
| Average = 96% | ||||||
| 2. | Undiluted | ---- | 5.66 | ---- | ||
| 1:2 | 2.83 | 2.86 | 101% | |||
| 1:4 | 1.42 | 1.55 | 109% | |||
| 1:8 | 0.71 | 0.67 | 94% | |||
Average = 101%
| ORIENT NEW LIFE MEDICAL CO., LTD. | |
| Contact: | Jerry Meng |
| Email: | Jerry @ newlifebiotest .com |
| Tel. | +86 18657312116 |
| SKYPE | enetjerry |