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Total Thyroxine (T4)
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Total Thyroxine (T4)
The Quantitative Determination of Total Thyroxine Concentration in Human Serum by a Chemiluminescence Immunoassay
Summary and explanation of the test
Measurement of the serum concentration of thyroxine (T4) is generally regarded as an important in-vitro diagnostic test for assessing thyroid function. This importance has provided the impetus for the significant improvement in assay methodology that has occurred in the last three decades. This microplate enzyme immunoassay methodology provides the technician with optimum sensitivity while requiring few technical manipulations. In this method, serum reference, patient specimen or control is first added to a microplate well. Enzyme-T4 conjugate is added, and then the reactants are mixed. A competition reaction results between the enzyme conjugate and the native thyroxine for a limited number of antibody combining sites immobilized on the well.
Materials provided for 96-well microplate
1. | Microplate | 1 piece | 96 test/piece |
2. | Calibrators | 6 tubes | 0.5 mL/tube |
Concentrate:0, 20, 40, 80, 160, 320ng/mL | |||
3. | Label | 1 bottle | 6 mL/bottle |
4. | Incubate buffer | 1 bottle | 6 mL/bottle |
5. | Substrate A Substrate B | 1 bottle 1 bottle | 6 mL/bottle 6 mL/bottle |
Before using, the substrates A and B are mixed by equal volume in practicality requisite amount. | |||
6. | Washing buffer | 1 bottle | 20 mL/bottle |
Before using, being diluted to 600ml with distilled water | |||
7. | Instruction book | 1 piece | 1 piece/kit |
Keeping in 2~8Degree Celsius , is 12 months.
Specimen collection and preparation
The specimens shall be blood serum in type and the usual precautions in the collection of venipuncture samples should be observed. For accurate comparison to established normal values, a fasting morning serum sample should be obtained. The blood should be collected in a plain red-top venipuncture tube with or without gel barrier. Allow the blood to clot. Centrifuge the specimen to separate the serum from the cells. Samples may be refrigerated at 2~8Degree Celsius for a maximum period of five days. If the specimen can not be assayed within this time, the sample may be stored at temperatures of -20Degree Celsius for up to 30 days. Avoid repetitive freezing and thawing.
Test procedure
The kit is in room temperature for more than 15mins, then
Unit:μL
Microplate Blank well Standards well Samples well |
Calibrators — 50 — Specimens — — 50 Incubate buffer — 50 50 Label — 50 50 |
Swirl the microplate gently for 2mins to mix and cover, incubate 45 mins at 37Degree Celsius . After washing for 5 times, the substrates are added. |
Substrate A 50 50 50 Substrate B 50 50 50 |
Or we also can mix the substrates in a bottle before using, and then pipette 100μL to each wells. |
Swirl and detect |
Detect After substrates being pipetted, suggest detecting in 1-3 mins by luminometer, one second per well. |
The data of concentrate and RLU is fitted in linear by log(X)- log(Y).
Expected values for the T4 (in ng/mL)
Low normal range 35
High normal range 140
Performance characteristics
A: Precision
With assay precision is less than 10%, between assay precision is less than 15%.
B: Accuracy
Between 90%-110%
C: Sensitivity
Less than 5ng/mL
Manufacturing enterprise: Beijing Savant Biotechnology Co., Ltd.
Registration: Beijing Drug Administration(prospective) 2400104, the first word in 2009
Total Thyroxine (T4)
