Document No. : INS-T4_F-EN (Rev.

05)
Revision date : December 22, 2017

Components of AFIAS T4
WARNINGS AND PRECAUTIONS Cartridge Box Contains
For in vitro diagnostic use only. - Cartridge 24
Carefully follow the instructions and procedures described in this ‘Instruction - Pipette Tip (Zipperbag) 24
for use’. - Spare Cartridge Zipperbag 1
Use only fresh samples and avoid direct sunlight. - ID Chip 1
Lot numbers of all the test components (cartridge and ID chip) must match - Instruction For Use 1
each other.

T4
INTENDED USE
AFIAS T4 is a fluorescence Immunoassay (FIA) for the quantitative determination
of total Thyroxine (total T4) in human serum/plasma. It is useful as an aid in
management and monitoring of thyroid disorders.
For in vitro diagnostic use only.
INTRODUCTION
Thyroxine (T4) is one of two major hormones produced by the thyroid gland (the
other is called triiodothyronine, or T3). T4 and T3 are regulated by a sensitive
feedback system involving the hypothalamus and the pituitary gland. The
hypothalamus releases the thyrotropin-releasing hormone (TRH), which stimulates
the pituitary to release the thyroid stimulating hormone (TSH). This causes the
thyroid to release T3 and T4 and these in turn regulate the release of TRH and TSH
via a feedback control mechanism. Normally, elevated blood levels of T4 and T3 act
to decrease the amount of TSH secreted, thereby reducing the production and
release of T4 and T3. Over 99 % of T4 is reversibly bound to three plasma proteins in
blood: thyroxine binding globulin (TBG) binds close to 70 %, thyroxine binding pre-
albumin (TBPA) binds 20 %, and albumin binds 10 %. Approximately 0.03 % of T4 is
in the free, unbound state in blood at any one time.
T4 is a useful marker for the diagnosis of hypothyroidism and hyperthyroidism. The
level of T4 decreases in hypothyroidism, myxedema and chronic thyroiditis
(Hashimoto’s disease). Increased levels of T4 have been found in hyperthyroidism
due to Grave’s disease and Plummer’s disease. The instrument for AFIAS tests T4
measures quantitatively T4 concentration in human serum/plasma.
PRINCIPLE
The test uses a competitive immunodetection method. In this method, the target
material in the sample binds to the fluorescence (FL)-labeled detection antibody in
detection buffer, to form the complex as sample mixture. This complex is loaded to
migrate onto the nitrocellulose matrix, where the covalent couple of T4 and bovine
serum albumin (BSA) is immobilized on a test strip, and interferes with the binding of
target material and FL-labeled antibody. If the more target material exists in blood,
the less detection antibody is accumulated, resulting in the less fluorescence signal.
COMPONENTS
AFIAS T4 consists of ‘Cartridge’, ‘Pipette tip’, ‘ID chip’ and ‘Instruction for use’.
Each cartridge packaged in an aluminum pouch has three components,
solution A part, solution B part and cartridge part.
Cartridge part contains a test strip, the membrane which has anti human T4-
BSA at the test line, while streptavidin at the control line.
Solution A part contains ANS and 0.1 % sodium azide as a preservative in
phosphate buffered saline (PBS).
Solution B part contains anti human T4-fluorescence conjugate, biotin-BSA-
fluorescence conjugate, bovine serum albumin (BSA) as a stabilizer and 0.1 %
sodium azide in phosphate buffered saline (PBS) as a preservative.
Do not interchange the test components between different lots or use the test
components after the expiration date, either of which might yield misleading
of test result(s).
The cartridge should remain sealed in its aluminum pouch until use. Do not
use the cartridge if that is damaged or already opened.
For shipping, samples must be packed in accordance with the regulations.
Sample with severe hemolytic and hyperlipidemia cannot be used and should
be recollected.
Just before use, allow the cartridge and sample to be at room temperature for
approximately 30 minutes.
AFIAS T4 as well as the instrument for AFIAS tests should be used away from
vibration and/or magnetic field. During normal usage, it can be noted that the
instrument for AFIAS tests may produce minor vibration.
Used pipette tips, and cartridges should be handled carefully and disposed by
an appropriate method in accordance with relevant local regulations.
An exposure to larger quantities of sodium azide may cause certain health
issues like convulsions, low blood pressure and heart rate, loss of
consciousness, lung injury and respiratory failure.
AFIAS T4 will provide accurate and reliable results subject to the following
conditions.
- AFIAS T4 should be used only in conjunction with the instrument for AFIAS
tests.
- Any anticoagulants other than sodium heparin, sodium citrate should be
avoided.
LIMITATIONS OF THE TEST SYSTEM
The test may yield false positive result(s) due to the cross-reactions and/or
non-specific adhesion of certain sample components to the capture/detector
antibodies.
The test may yield false negative result. The non-responsiveness of the
antigen to the antibodies is most common where the epitope is masked by
some unknown components, so as not to be detected or captured by the
antibodies. The instability or degradation of the antigen with time and/or
temperature may cause the false negative as it makes antigen unrecognizable
by the antibodies.
Other factors may interfere with the test and cause erroneous results, such as
technical/procedural errors, degradation of the test components/reagents or
presence of interfering substances in the test samples.
Any clinical diagnosis based on the test result must be supported by a
comprehensive judgment of the concerned physician including clinical
symptoms and other relevant test results.
STORAGE AND STABILITY
The cartridge is stable for 20 months (while sealed in an aluminum pouch) if
stored at 2-8 °C.
Return an unused cartridge to the spare cartridge zipperbag containing the
desiccant pack. Reseal along entire edge of zip-seal. May be stored for up to 1
month at 2-8℃.
MATERIALS SUPPLIED
REF SMFP-19
MATERIALS REQUIRED BUT SUPPLIED ON DEMAND
Following items can be purchased separately from AFIAS T4.
Please contact our sales division for more information.
AFIAS-1 REF FPRR019
AFIAS-6 REF FPRR020
Boditech Hormone Control REF CFPO-95
Boditech Hormone Calibrator REF CFPO-107
SAMPLE COLLECTION AND PROCESSING
The sample type for AFIAS T4 is human serum/plasma.
It is recommended to test the sample within 24 hours after collection.
The serum or plasma should be separated from the clot by centrifugation
within 3 hours after the collection of whole blood.
Samples may be stored for up to a week at 2-8 °C prior to being tested. If
testing will be delayed more than a week, samples should be frozen at -20 °C
or below.
Samples stored frozen at -20 °C for 3 months showed no performance
difference.
Once the sample was frozen, it should be thawed one time and only for test,
because repeated freezing and thawing can result in the changed test values.
Sample must be mixed thoroughly after by low speed vortexing or by gently
inverting, and centrifuged prior to use to remove particulate matter and to
ensure consistency in the results.
TEST SETUP
Check the components of the AFIAS T4 as described below. : Cartridge, pipette
tip, ID chip and instruction for use
Keep the sealed cartridge (if stored in refrigerator) at room temperature for
at least 30 minutes just prior to the test. Place the cartridge on a clean, dust-
free and flat surface.
Turn on the instrument for AFIAS tests.
Empty the tip box.
Insert the ID chip into the “ID chip port”.
Please refer to the instrument for AFIAS tests ‘Operation Manual’ for complete
information and operating instructions.
TEST PROCEDURE
1) Take 150 µL of sample with a pipette and dispense it into the sample well
on the cartridge.
2) Insert the cartridge into the cartridge holder.
3) Insert a tip into the tip hole of the cartridge.
4) Tap the ‘START’ icon on the screen.
5) The test result will be displayed on the screen after 10 minutes.
※ Note: Refer to the instrument for AFIAS tests Operation Manual to select
sample type.
INTERPRETATION OF TEST RESULT
The instrument for AFIAS tests calculate the test result automatically and
displays T4 concentration of the test sample in terms of nmol/L.
Document No. : INS-T4_F-EN (Rev. 05)
Revision date : December 22, 2017

The reference range*: 57.9-150.6 nmol/L [nmol/L] Avg. CV% Avg. CV% Avg. CV% Avg. CV% Avg. CV%
The working range of the AFIAS T4 is 10.23-300 nmol/L. 60 62.18 3.2 61.61 4.4 61.04 3.7 61.61 4.4 61.41 3.8
Conversion factor: 12.87 (SI: nmol/L = 12.87 x μg/dL) 120 119.77 4.8 118.66 3.7 121.30 3.3 118.66 3.7 121.05 3.7
*National institutes of health. 180 181.62 2.1 180.63 1.1 181.01 1.3 180.63 1.1 180.18 2.1
Accuracy Note: Please refer to the table below to identify various symbols.
QUALITY CONTROL The accuracy was confirmed by 3 different lots testing six times each different
Quality control tests are a part of the good testing practice to confirm the concentrations.
expected results and validity of the assay and should be performed at regular Expected value [nmol/L] Lot 1 Lot 2. Lot 3 AV Recovery (%)
intervals. 15.22 15.22 15.20 15.17 15.20 99.8
The control tests should be performed immediately after opening a new test 72.88 72.88 75.81 78.12 75.60 103.7
lot to ensure the test performance is not altered. 103.06 103.06 102.83 102.03 102.64 99.6
Quality control tests should also be performed whenever there is any question 144.65 144.65 149.41 152.83 148.97 103.0
concerning the validity of the test results. 249.88 249.88 251.87 249.96 250.57 100.3
Control materials are not provided with AFIAS T4. For more information
Comparability
regarding obtaining the control materials, contact Boditech Med Inc.’s Sales
Division for assistance. T4 concentrations of 81 serum samples were quantified independently
(Please refer to the instruction for use of control material.) with AFIAS T4 and mini VIDAS®(BioMerieux Inc. France) as per prescribed
test procedures. Test results were compared and their comparability was
investigated with linear regression and coefficient of correlation (R).
PERFORMANCE CHARACTERISTICS
Linear regression and coefficient of correlation between the two tests
Analytical Sensitivity were Y=0.9726X – 13.594 and R = 0.9654 respectively.
Limit of Blank (LoB) 0.37 nmol/L
Limit of Detection (LoD) 2.36 nmol/L
Limit of Quantification (LoQ) 10.23 nmol/L
Analytical Specificity
- Cross reactivity
There was no significant cross-reactivity from these materials with the AFIAS
T4 test measurements.
Cross reactants (Conc.) Cross reactivity (%)
l-Triiodothyronine (500 ng/mL) 3.2
reverse T3 (500 ng/mL) 3.6
l-Thyrosine (300 ng/mL) 0.7
d-Thyrosine (300 ng/mL) 1.2 For technical assistance; please contact:
3-lodo-L-tyrosine (500 ng/mL) 0.9 Boditech Med Inc.’s Technical Services
salicylic acid (1,000,000 ng/mL) ND Tel: +82 33 243-1400
*ND: Not Detected E-mail: sales@boditech.co.kr

- Interference
Study of interference from table below with AFIAS T4 showed following Boditech Med Incorporated
results. Cholesterol had effect on AFIAS T4 test in the procedure. 43, Geodudanji 1-gil, Dongnae-myeon,
Interference materials (Conc.) Interference (%) Chuncheon-si, Gang-won-do, 24398
D-glucose (60 mM/L) 3.5 REFERENCES Republic of Korea
L-Ascorbic acid (0.2 mM/L) 4.0 1. Thakur C., Saikia T.C. Yadav R.N., Total serum levels of triiodothyronin(T3) Tel: +(82) -33-243-1400
Bilirubin (0.4 mM/L) 2.7 thyroxine(T4) and thyrotropine(TSH) in school going children of Dibrugarh district: Fax: +(82) -33-243-9373
Hemoglobin (2 g/L) 4.8 an endemic goiter region of Assam. Indian J Physiol Pharmacol, 1997, 41(2) : 167- www.boditech.co.kr
170
Cholesterol (13 mM/L) 6.3 2. Larsen P.R., Dockalova J., Sipula D., Wu F. M. Immunoassay of Thyroxine in Obelis s.a
Triglyceride (10 mg/ml) 3.0 unextracted Human Serum. J. Clin. Endocrinol. Metabl., 1973, 37(2):177-182
3. Wagner M. S., Wajner S. M., Maia A. L. The Role of Thyroid Hormone in testicular Bd. Général Wahis 53,
Precision Development and Function. J. Endocrinol., 2008, 199(3) : 351-365 1030 Brussels, BELGIUM
[Intra-assay] The intra-assay precision was calculated by one evaluator, who 4. Wahlin A., Wahlin T. B., Small B. J., Backman L. Influences of thyroid stimulating Tel: +(32) -2-732-59-54
tested different concentration of control standard 20 times each with three hormone on cognitive functioning in very old age. J. Gerontol B. Psychol Sci. Soc. Fax: +(32) -2-732-60-03
Sci., 1998, 5 : 234-239 E-Mail: mail@obelis.net
different lots of AFIAS T4. 5. Willian R. Kirk, et al. Characterization of the sources of protein-Ligand Affinity:1-
T4 [nmol/L] Lot 1 Lot 2 Lot 3 Avg. SD CV% sulfonato-8-(1’) anilinonaphthalene Binding to intestinal Fatty Acid binding protein.
60 61.75 61.99 62.79 62.18 2.01 3.2 Biophysical Jounal(1996) pp.69-83
120 118.40 120.87 120.05 119.77 5.76 4.8 6. Jeramia K. Ory and Leonard J. Banaszak. Studies of the Binding Reaction of
180 181.07 182.35 181.44 181.62 3.77 2.1 Adipocyte Lipid Binding Protein using the Fluorescent Probe 1,8-
Anilinonaphthalene-8-Sulfonate. Biophysical Journal(1999) pp.1107-1116
[Inter-assay] The inter-assay precision was confirmed by 3 different evaluators 7. Toshiro Sakurada, et al. Radioimmunoassay for serum Thyroxine. Tohoju J.(1976)
with 3 different lot AFIAS T4 during 5 days testing 3 times each different pp.125-133
concentration.
T4 Between lot Between person Between day Between site Total