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)