BodiTech Med Inc. i-CHROMATMT4

i-CHROMATMT4

ImmunoAssay for Quantitative Measurement of T4 in Human Serum and Plasma with thei-CHROMATM Readersystem.

INTENDED USE

i-CHROMATMT4along with i-CHROMATMReader is a fluorescence immunoassay that quantifies the concentration of T4in human serum and plasma.

INTRODUCTION

Thyroxine (T4) is one of two major hormones produced by the thyroid gland (the other is called triiodothyronin, or T3). T4 and T3 are regulated by a sensitive feedback system involving the hypothalamus and thepituitary 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.i-CHROMATMT4 measuresquantitativelyT4 concentration inhuman serum/plasma.

PRINCIPLE

i-CHROMATMT4uses a competitive immunodetection method, such that the detector antibody in buffer binds to T4 in the bloodsample and antigen-antibody complexes competes with T4 covalently coupled to BSA that has been immobilized on test strip as sample mixture migrates through the nitrocellulose matrix. Thus the more T4 antigen in blood, the less antigen-antibody complexes accumulated on the test strip. The fluorescence intensity of the detector antibody reflects the amount of antigen captured and is processed in i-CHROMATMReader to determine the T4 concentration in the specimen. The working range ofi-CHROMATMT4 is 20~300 nmol/L.

* Normal Value : 60 ~ 160 nmol/L

MATERIALS PROVIDED

BodiTech Med Incorporated i-CHROMATMT4

Catalog REF Catalog No. 13018

kit contains:

Test device 25T/box

Detection buffer 25 tubes/pouch

ID Chip 1ea/box

Insert 1 sheet/box

MATERIALSREQUIRED BUT NOT PROVIDED

i-CHROMATM Reader REF Catalog No.FR-203

ReceiptPrinter

Transfer pipette (10µL,75µL size)

COMPOSITION OF REAGENTS

i-CHROMATMT4 consists of a test device, a detection buffer andan ID chip. The test device is individually sealed with a desiccant in an aluminum pouch, and the detection buffer is dispensed individually in a tube. A pouch containing the pre-dispensed tubes is delivered separately from the test device in a styrofoam box filled with ice packs.

•Test devicecontains a test strip in which BSA-labeledT4 and streptavidin have been immobilized on the test and the control line of striprespectively.

•Detection buffer, predispensed in a tube, contains fluorescence-labeled anti-T4 antibody, fluorescence labeled biotin-BSA, gelatin as a stabilizer, and sodium azide as a preservative in PBS.

WARNINGS AND PRECAUTIONS

• IVD For In Vitro Diagnostic Use.

•Carefully follow the instructions and procedures described in this insert. REF Catalog No.13018

• Don’t use the test device if its lot # does not match with that on the ID chip.

•i-CHROMATMT4works only in i-CHROMATMReader. Andtests should be performed by trainedstaffsfromthe laboratories where the sample(s) is taken byqualified medical personnel.

•LOTNeither interchange materials from different product lots nor use beyond the expiration date. The use of the medical test device beyond expiration date may affect on test result.

•Thei-CHROMATMT4test device should remain in itsoriginal sealed pouch until ready to use. Do notuse test deviceif the pouch is damaged or the seal is broken. Discardafter single use.

•Thei-CHROMATMT4test deviceand the reader should be used away from vibration and magnetic field. During the normal usage, i-CHROMATMT4 may introduce minute vibration, which should be regarded normal.

•Use separate clean pipette tipsand detection buffer tubefordifferent specimens.The pipette tips and detection buffer tubeshould be used for one specimen only. Discardafter single use.

• Blood specimens, used test devices, pipette tips and detection buffer tube arepotentially infectious. Proper laboratory safety techniques, handling and disposalmethods shouldbe followed in accordance with standard procedures and relevant regulations observed by microbiological hazard materials.

•The test will be applied on a routine basis and not in emergencysituations.

•Do not smoke, eat, or drink in areas in which specimens or kit reagents are handled.

STROAGE AND STABILITY

•Store the detection bufferin a refrigerator at 2° - 8°C. The Detection buffer is stable up to 20 months if stored in a refrigerator.

•Once removed from refrigerator, allow the detection buffer for 20 minutes to warm up to the room temperature before testing.

• Storethetest devices at 4°-30°C in the sealed pouch they came in.Thetest deviceis stable for 20 months (while in the sealed pouch)ifstored at 4°-30°C.

•If stored in a refrigerator, allow a minimum of 20 minutesfor the test device and detection buffer to reach room temperature while it is in the sealedpouch.

•Do not remove the test device from the pouch until ready to use.Thetest device should be used immediately once opened.

• The storage and shipping of test kit should be complied as indicated in the instruction manual. However, it is remotely possible that only part of test kit is affected bystability problems.

SAMPLE COLLECTIONANDPREPARATION

The test can be performed with either serum or plasma.

•For the serum sample, collect the blood in a tube without anticoagulant and allow to be clotted. Remove the serum from the clot as soon as possible to avoid hemolysis. For the plasma sample, collect the blood in a tube treated with EDTA. Anticoagulants other than EDTA for plasma specimen have not been evaluated. If testing cannot be conducted within an hour after preparation of specimen, the serum/plasma should be stored at -20o C until tested. When performing tests, apply it immediately after specimen was taken.

•The specimen must be at room temperature and be homogeneous before testing. Frozen specimens must be completely thawed, thoroughly mixed, and brought to room temperature prior to testing. If specimens are to be shipped, they should be packed in compliance with regulations.

•It is recommended to avoid using severely hemolyzed specimens whenever possible. If a specimen appears to be severely hemolyzed, another specimen should be obtained and tested.

ASSAY PROCEDURE

  1. Set a test device on a dust-free clean place.
  2. Check/insert ID Chip onto the instrument. Make sure that the test device lot # matches with that on the ID Chip.
  3. Take out one tube of the detection bufferfrom refrigerator and leave it at room temperature for 20 minutes or longer.
  4. Draw 10µL of serum/plasma or control with a transfer pipette and add it to the tube containing the detection buffer.
  5. Mix well the specimen with the detection buffer by tapping or inverting the tube.
  6. Take 75 µL of sample mixture and load it onto the well of the test device.
  7. Leave the test device at the room temperature for 10min before inserting the test device into the reader.
  8. Insert the test deviceonto the holder of thei-CHROMATM Readerandpress the“SELECT”key. Read the results on the display screen of i-CHROMATMReader.

Refer toi-CHROMATMReader operationmanual for the complete instruction on the use of the test.

RESULT

i-CHROMATMReaderfinds and displays the T4valueon the LC display in units of nmol/L.For further information, refer to the operation manual fori-CHROMATM Reader.

QUALITY CONTROL

A quality control test should be performed as a part of the good testing practice, to confirm the expected QC results, to confirm the validity of the assay, and to assure the accuracy of patient results. QC specimens should also be run whenever there rises any question concerning the validity of results obtained. Upon confirmation of the expected result, the test deviceis ready to use with patient specimens. Control standards are not provided with this test kit. If QC of the test kit should be desired, Lyphochek® Immunoassay Plus Control is recommended by Boditech Med Inc.

i-CHROMATMT4contains internal procedure control thatsatisfies routine quality control requirements. Thisinternal control is performed each time a patient sample is tested. This controlindicates that the The test device was properly inserted and read by i-CHROMATM Reader.An invalid result from the internal control causes anerror message on i-CHROMATM Reader indicating that the test should berepeated.

LIMITATIONS OF THE PROCEDURE

•The result of i-CHROMATMT4 should be evaluated together with all clinicaland laboratory data available. If T4 test results do not agree with the clinical evaluation, additional tests should be performed.

• The false positive resultincludes cross-reactions withsome components of serum from individuals to antibodies; and non-specific adhesionof some components in human blood that have similar epitopes to capture and detector antibodies. In the case of false negativeresults, the most common factors are: non-responsiveness of antigen to the antibodies by that certain unknown components are masking its epitope, such that antigen cannot be seen by the antibodies; instability of T4antigen,resulting in degradation with time and, or temperature, such that they become no longerrecognizable by antibodies, anddegraded other degraded test components. The effectiveness of thetestis highlydependent on storage of kits and sample specimens at optimal conditions.

• Plasma using anticoagulants (e.g. heparin or citrate) other than EDTA has not been evaluated in i-CHROMATMT4and thus should not be used.

•Other factors may interfere with i-CHROMA TMT4and may causeerroneous results. These include technical or procedural errors, as well asadditional substances in blood specimens.

PERFORMANCE CHRACTERISTICS

  1. Analytical Sensitivity

i-CHROMATMT4was evaluated to determine the limit of detection. Three different lots of test devices were evaluated with 10 runsof each lot. The minimum detection limit was defined as +3SD over the average result from null specimens. The limit of i-CHROMATMT4was found to be 6.76nmol/L .

  1. Specificity

The following compounds were tested at concentrations up to1,000nmol/L for cross-reactivity:There was no significant interference withT4measurement, nor was there any significant assay cross-reactivity with those bio-molecules tested.

Compound / Spiked concentration(nmol/L) / %Cross-reactivity
Tetraiodothyroacetic Acid / 300 / 5.16
Monoiodotyrosine / 1,000 / 0.01
Diiodotyrosine / 1,000 / <0.01
Phenytoin / 1,000 / <0.01
Phenylbutazone / 1,000 / <0.01
l-Triiodothyronine / 500 / 1.24
d-Triiodothyronine / 500 / 0.81
l-Tyrosine / 5,000 / <0.01
d-Tyrosine / 5,000 / <0.01

3.Precision: For the intra-assay imprecision, 20 replicates were tested at each control sample. For the inter-assay imprecision, tests were conducted on 10 sequential days, with 10 runs per day and with 10 replicates at each T4 concentration.

Intra- and inter-assay precision
Intra-assay / Inter-assay
Sample / Mean / CV / Mean / CV
(nmol/L) / (nmol/L) / (%) / (nmol/L) / (%)
31.2 / 31.5 / 4.6 / 31.2 / 4.9
79.4 / 79.9 / 5.0 / 80.5 / 4.3
118.5 / 121.1 / 4.1 / 121.5 / 4.4
285.2 / 289.3 / 1.5 / 289.6 / 2.2

4.Comparability

T4 concentrations of 103clinical specimens were quantified independently with i-CHROMATMT4and the Beckmen Coulter Access2 Immunoassay system according to the established standard test procedure. The test results wereanalyzed and their compatibilities were investigated with linear regression and correlation coefficient (R). The value of correlation of coefficient was 0.916 between two methods.

REFERENCES

1. Thakur C., Saikia T.C. Yadav R.N., Total serum levels of triiodothyronin(T3) thyroxine(T4) and thyrotropine(TSH) in school going children of Dibrugarh district: an endemic goiter region of Assam. Indian J Physiol Pharmacol, 1997, 41(2) : 167-170

2. Larsen P.R., Dockalova J., Sipula D., Wu F. M. Immunoassay of Thyroxine in 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 Development and Function. J. Endocrinol., 2008, 199(3) : 351~365

4. Wahlin A., Wahlin T. B., Small B. J., Backman L. Influences of thyroid stimulating hormone on cognitive functioning in very old age. J. Gerontol B. Psychol Sci. Soc. Sci., 1998, 5 : 234~239

BodiTech Med’sexpress and implied warranties (including impliedwarranties of merchantability and fitness) are conditional uponobservance of BodiTech Med’s published directions with respect to the useof BodiTech Med’s products.

For Technical Assistance call

Boditech Med’s Technical Services at

Tel: +82 (33) 243-1400

E-mail:

BodiTech Med Incorporated

G-Tech Village, 1144-2 Geoduri Dongnaemyeon,

Chuncheon, Gangwondo200-883

Republic of Korea

Tel: +82 (33) 243-1400, Fax: +82 (33) 243-9373

E-Mail:

EU Representative:Jai Jun Choung, Ph.D.

Eu Biotech Development Ltd.

81 Oxford Street, London, W1D 2EU
United Kingdom
Tel: +44 207 903 5441, Fax: +44 207 903 5333

E-Mail:

i-CHROMATM is registered trademarks of BodiTech MedIncorporated.

Revision No: 00

Date of last revision: November 01, 2010.

BodiTech Med Inc.

Rev. 00_101101_M Enjoy Your Life!