30 441 09083 G - en - 2015/06

VIDAS® TSH3 (TSH3)

VIDAS TSH3 is an automated quantitative ultra-sensitive test for use on the VIDAS family instruments, for the third
generation immunoassay of human thyrotropin hormone in human serum or plasma (siliconized tube, plain tube with
separator gel or tube with lithium heparin) using the ELFA technique (Enzyme Linked Fluorescent Assay).

SUMMARY AND EXPLANATION PRINCIPLE

VIDAS TSH3 improves the sensitivity and accuracy of The assay principle combines a one-step enzyme
the TSH assay. immunoassay sandwich method with a final fluorescent
Thyrotropin or thyroid-stimulating hormone (TSH) is a detection (ELFA).
®
glycoprotein with a molecular weight of 28,000 to The Solid Phase Receptacle (SPR ) serves as the solid
30,000 daltons. TSH is composed of two non-covalently phase as well as the pipetting device for the assay.
bound α and β peptide subunits. The α subunit, Reagents for the assay are ready-to-use and pre-
composed of 92 amino acids, is similar to that of FSH, LH, dispensed in the sealed reagent strips.
and hCG. The β subunit, composed of 112 amino acids, All of the assay steps are performed automatically by the
determines the specific biological and immunological instrument. The reaction medium is cycled in and out of
properties of the hormone. Glycan residues are bound to the SPR several times.
these subunits (1,4). The sample is transferred into the well containing anti-
TSH is produced by thyrotropic cells in the anterior TSH antibody labeled with alkaline phosphatase
pituitary gland. It is secreted into the bloodstream (conjugate). The sample/conjugate mixture is cycled in
according to a circadian rhythm, peaking at between and out of the SPR. The antigen binds to antibodies
1 and 2 a.m. coated on the SPR and to the conjugate forming a
TSH stimulates the thyroid gland to produce the main "sandwich". Unbound components are eliminated during
thyroid hormones T3 and T4. In return, these thyroid the washing steps.
hormones exert a negative-feedback effect on the pituitary Two final detection steps are successively performed. At
gland, reducing TSH secretion. TSH secretion is also each step, the substrate (4-Methyl-umbelliferyl phosphate)
influenced by the central nervous system, via a is cycled in and out of the SPR. The conjugate enzyme
hypothalamic neuropeptide, TRH, and neuromediators catalyzes the hydrolysis of the substrate into a fluorescent
such as somatostatin or dopamine. product (4-Methyl-umbelliferone), the fluorescence of
In cases of hyperthyroidism (Basedow's disease, thyroid which is measured at 450 nm. The intensity of the
adenoma, and inflammatory thyroiditis), TSH level is fluorescence is proportional to the concentration of
severely inhibited and may even be undetectable. In rare antigen present in the sample.
forms of high-origin hyperthyroidism, the TSH level is not At the end of the assay, results are automatically
reduced, since the negative-feedback control of the calculated by the instrument in relation to two calibration
thyroid hormones has no effect. curves stored in memory corresponding to the two
In cases of clear-cut primary hypothyroidism, TSH levels detection steps. A threshold signal determines the choice
are always much higher than normal and thyroid hormone of the calibration curve to be used for each sample. The
levels are low. results are then printed.
In partial or mild hypothyroidism, a moderate increase in
the TSH level enables normal thyroid production to be
maintained for many years without any apparent clinical
symptoms (2).
The VIDAS TSH3 assay aids in diagnosing thyroid or
hypophysial disorders.

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CONTENT OF THE KIT (60 TESTS) – RECONSTITUTION OF REAGENTS:

60 TSH3 strips STR Ready-to-use.
60 TSH3 SPRs SPR Ready-to-use.
2 x 30 SPRs sensitized with monoclonal anti-TSH immunoglobulins (mouse).
TSH3 controls: Reconstitute with 2 ml of distilled water. Wait for 5 to 10 minutes. Mix. Stable after
C1 control C1 reconstitution for 14 days at 2-8°C or until the expiration date on the kit at
1 x 2 ml (lyophilized) - 25 ± 6°C. Five freeze/thaw cycles are possible.
C2 control Human serum* + human TSH + preservatives.
1 x 2 ml (lyophilized) C2 MLE data indicate the confidence interval in “µIU/mL” (micro-international units
per milliliter) ("Control C2 Dose Value Range").
TSH3 calibrators: Reconstitute with 3 ml of distilled water. Wait for 5 to 10 minutes. Mix. Stable after
S1 calibrator S1 reconstitution for 14 days at 2-8°C or until the expiration date on the kit at - 25 ±
1 x 3 ml (lyophilized) 6°C. Five freeze/thaw cycles are possible.
S2 calibrator Processed calf serum + human TSH + preservatives.
1 x 3 ml (lyophilized) S2 MLE data indicate the concentration in µIU/mL ("Calibrator (S2) Dose Value") and
the confidence interval in "Relative Fluorescence Value" ("Calibrator (S2) RFV
Range").
TSH3 diluent R1 Ready-to-use.
1 x 2 ml (liquid) Processed calf serum + 0.9 g/l sodium azide.
Specifications for the factory master data required to calibrate the test:
• MLE data (Master Lot Entry) provided in the kit,
or
• MLE bar code printed on the box label.
1 Package insert provided in the kit or downloadable from www.biomerieux.com/techlib.
* This product has been tested and shown to be negative for HBs antigen, antibodies to HIV1, HIV2 and HCV. However, since no
existing test method can totally guarantee their absence, this product must be treated as potentially infectious. Therefore, usual safety
procedures should be observed when handling.

The SPR The strip

The SPR is coated during production with anti-TSH The strip consists of 10 wells covered with a labeled, foil
immunoglobulins (mouse). Each SPR is identified by the seal. The label comprises a bar code which mainly
TSH3 code. Only remove the required number of SPRs indicates the assay code, kit lot number and expiration
from the pouch and carefully reseal the pouch after date. The foil of the first well is perforated to facilitate the
opening. introduction of the sample. The last well of each strip is a
cuvette in which the fluorometric reading is performed.
The wells in the center section of the strip contain the
various reagents required for the assay.
Description of the TSH3 strip
Wells Reagents
1 Sample well.
2-3-4 Empty wells.
5 Conjugate: alkaline phosphatase-labeled monoclonal anti-TSH immunoglobulins (mouse)
+ 0.9 g/l sodium azide (400 µl).
6-7-8-9 Wash buffer: Tris (0.01 mol/l, pH 7.6) + NaCl (0.15 mol/l) + Tween 1% + 0.9 g/l sodium azide (600 µl).
10 Cuvette with substrate: 4-Methyl-umbelliferyl-phosphate (0.6 mmol/l)
+ diethanolamine*(0.62 mol/l or 6.6 %, pH 9.2) + 1 g/l sodium azide (300 µl).

* Signal Word: DANGER

Hazard statement
H318 : Causes serious eye damage.
Precautionary statement
P280 :Wear protective gloves/protective clothing/eye protection/face protection.
P305 + P351 + P338: IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if present
and easy to do. Continue rinsing.
For further information, refer to the Material Safety Data Sheet.

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MATERIALS AND DISPOSABLES REQUIRED BUT STORAGE CONDITIONS

NOT PROVIDED • Store the VIDAS TSH3 kit at 2-8°C.
- Pipette with disposable tip to dispense 2 ml, 3 ml and • Do not freeze the SPRs and strips.
200 µl. • Store all unused reagents at 2-8°C.
- Powderless, disposable gloves. • After opening the kit, check that the SPR pouch is
- For other specific materials and disposables, please correctly sealed and undamaged. If not, do not use the
refer to the Instrument User’s Manual. SPRs.
- VIDAS family instrument. • Carefully reseal the pouch with the desiccant inside
after use to maintain stability of the SPRs and return
WARNINGS AND PRECAUTIONS the complete kit to 2-8°C.
• For in vitro diagnostic use only. • If stored according to the recommended conditions, all
• For professional use only. components are stable until the expiration date indicated
• This kit contains products of human origin. No on the label. Refer to the kit composition table for
known analysis method can totally guarantee the special storage conditions.
absence of transmissible pathogenic agents. It is
SPECIMENS
therefore recommended that these products be
treated as potentially infectious and handled Specimen type and collection:
observing the usual safety precautions (see Serum or plasma (collected in a siliconized tube, plain
Laboratory Biosafety Manual - WHO - Geneva - tube with separator gel or tube with lithium heparin).
latest edition). It is recommended that each laboratory checks the
• This kit contains products of animal origin. Certified compatibility of collection tubes used.
knowledge of the origin and/or sanitary state of the Do not use samples collected using plain tubes with
animals does not totally guarantee the absence of glycerol-coated stoppers.
transmissible pathogenic agents. It is therefore Do not use plasma collected on EDTA, since the
recommended that these products be treated as presence of EDTA can lead to reduced measurement
potentially infectious and handled observing the usual values.
safety precautions (do not ingest or inhale).
None of the following factors have been found to
• Do not use the SPRs if the pouch is pierced.
significantly influence this assay.
• Do not use visibly deteriorated STRs (damaged foil or
- hemolysis (after spiking samples with hemoglobin:
plastic).
0 to 300 µmol/l (monomer)),
• Do not use reagents after the expiration date indicated
- lipemia (after spiking samples with lipids:
on the label.
0 to 11.4 mmol/l equivalent in triglycerides),
• Do not mix reagents (or disposables) from different lots.
- bilirubinemia (after spiking samples with bilirubin:
• Use powderless gloves, as powder has been reported
0 to 326 µmol/l).
to cause false results for certain enzyme immunoassay
However, it is recommended not to use samples that are
tests.
clearly hemolyzed, lipemic or icteric and, if possible, to
• Kit reagents contain sodium azide which can react with
collect a new sample.
lead or copper plumbing to form explosive metal azides.
If any liquid containing sodium azide is disposed of in Specimen stability:
the plumbing system, drains should be flushed with Samples can be stored at 2-8°C in stoppered tubes for up
water to avoid build-up. to 48 hours; if longer storage is required, freeze the sera
• The optical cuvette with substrate (well 10) contains an or plasma at - 25 ± 6°C. Avoid successive freezing and
irritant agent (diethanolamine). Refer to the hazard thawing.
statements "H" and the precautionary statements "P"
above. INSTRUCTIONS FOR USE
• Spills should be wiped up thoroughly after treatment For complete instructions, see the User’s Manual.
with liquid detergent and a solution of household bleach
containing at least 0.5% sodium hypochlorite. See the Reading Master lot data
User’s Manual for cleaning spills on or in the instrument. Before each new lot of reagents is used, enter the
Do not autoclave solutions containing bleach. specifications (or factory master data) into the instrument
• The instrument should be regularly cleaned and using the master lot entry (MLE) data.
decontaminated (see the User’s Manual).
• To obtain the performance described, strictly If this operation is not performed before initiating the
comply with the following precautions: tests, the instrument will not be able to print results.
- wear powderless gloves to handle SPRs and Note: the master lot data need only be entered once
strips, for each lot.
- remove each SPR from the pouch without It is possible to enter MLE data manually or
touching the tip, automatically depending on the instrument (refer to the
User’s Manual).
- manipulate each strip using its handle to
completely avoid touching the aluminium foil and
reading cuvette.

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Calibration QUALITY CONTROL

Calibration, using the two calibrators provided in the kit, Two controls are included in the VIDAS TSH3 kit. These
must be performed upon receipt of a new lot of reagents controls must be performed immediately after opening a
after the master lot data have been entered. Calibration new kit to ensure that reagent performance has not been
should then be performed every 14 days. This operation altered. Each calibration must also be checked using this
provides instrument-specific calibration curves and control. The instrument will only be able to check the
compensates for possible minor variations in assay signal control values if they are identified by C1 and C2.
throughout the shelf-life of the kit. The calibrators, Results cannot be validated if the control values deviate
identified by S1 and S2, must be tested in duplicate from the expected values.
(see User’s Manual).
The calibrator values must be within the set RFV "Relative Note
Fluorescence Value" range. If this is not the case, It is the responsibility of the user to perform Quality
recalibrate with S1 and S2. Control in accordance with any local applicable
regulations.
Procedure
LIMITATIONS OF THE METHOD
1. Only remove the required reagents from the
refrigerator and allow them to come to room Interference may be encountered with certain sera
temperature for at least 30 minutes. containing antibodies directed against reagent
2. Use one "TSH3" strip and one "TSH3" SPR for each components.
sample, control or calibrator to be tested. Make sure The results of a TSH3 assay must be interpreted as part
the storage pouch has been carefully resealed of a complete clinical profile. In case of discrepancy, this
after the required SPRs have been removed. assessment must be completed by thyroid hormone
3. The test is identified by the "TSH3" code on the measurement.
instrument. The calibrators must be identified by "S1"
and "S2", and tested in duplicate. If the controls are RANGE OF EXPECTED VALUES
to be tested, they should be identified by "C1" and The study involved 76 hyperthyroid patients, 11 patients
"C2". with severe non-thyroid pathologies, 180 euthyroid
4. Mix the calibrators, controls and samples using a patients and 47 patients with primary hypothyroidism.
vortex-type mixer (for serum or plasma separated from The thresholds obtained by ROC curve analysis (5)
the pellet). (NCCLS GP10-A standards) are as follows:
5. For this test, the calibrator, control, and sample Hyperthyroidism-euthyroidism threshold = 0.27 µIU/ml.
test portion is 200 µl. (sensitivity 100%; specificity 95.3%)
6. Insert the "TSH3" SPRs and "TSH3" strips into the Euthyroidism-primary hypothyroidism threshold = 4.7 µIU/ml.
instrument. Check to make sure the color labels with (sensitivity 93.8%; specificity 100%)
the assay code on the SPRs and the Reagent Strips
match. These figures are given as a guide; it is recommended
that each laboratory establish its own reference values
7. Initiate the assay as directed in the User’s Manual. All from a rigorously selected population.
the assay steps are performed automatically by the
instrument. PERFORMANCE
8. Reclose the vials and return them to the required Studies performed using VIDAS TSH3 gave the following
temperature after pipetting. results:
9. The assay will be completed within approximately 80
minutes. After the assay is completed, remove the Measurement range
SPRs and strips from the instrument. The measurement range of the VIDAS TSH3 kit extends
10. Dispose of the used SPRs and strips into an up to 100 µIU/ml.
appropriate recipient. Analytical detection limit
RESULTS AND INTERPRETATION Defined as the smallest concentration of TSH which is
significantly different from the zero concentration with a
Once the assay is completed, results are analyzed
probability of 95%: 0.005 µIU/ml.
automatically by the computer.
The results are automatically calculated by the instrument Functional detection limit
using two calibration curves which are stored by the The TSH concentration measured with an inter-assay
instrument (4 parameter logistic model) and are variation coefficient of 20% is less than 0.02 µIU/ml.
expressed in µIU/mI (2nd IRP 80/558). Samples with TSH The VIDAS TSH3 kit is denominated as a third generation
concentrations greater than 100 µIU/ml must be assay (3).
reassayed after dilution in TSH3 diluent (R1). If the
dilution factor has not been entered when the Work List Hook effect
was created (see User’s Manual), multiply the result by No hook effect was found up to TSH concentrations of
the dilution factor to obtain the sample concentration. 2500 µIU/ml.
Interpretation of test results should be made taking into
consideration the patient history, and the results of any
other tests performed.

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Precision
Six samples were tested in duplicate in 15 different runs (one run per day) with two reagent lots (N=60) on the same
VIDAS.
Using this protocol, intra-assay precision (within-run reproducibility), inter-assay precision (between-run reproducibility)
and total precision (inter-assay and inter-lot reproducibility) were calculated.

Intra-assay precision Inter-assay precision Total precision

Sample Mean Standard CV Standard CV Standard CV

µIU/ml deviation % deviation % deviation %
µIU/ml µIU/ml µIU/ml
1 0.019 0.0007 3.63 0.0013 6.79 0.0016 8.30
2 0.195 0.0037 1.91 0.0063 3.22 0.0110 5.65
3 0.995 0.0224 2.25 0.0437 4.40 0.0471 4.74
4 4.56 0.066 1.46 0.168 3.70 0.286 6.26
5 35.17 0.897 2.55 1.336 3.80 2.367 6.73
6 62.24 1.754 2.82 2.400 3.86 5.691 9.14

Specificity
Tested compounds Cross-reactivity (%)
TSH 100
LH < 0.01
FSH < 0.01
hCG < 0.01

Accuracy

Dilution test
Three samples were diluted in the TSH3 diluent and tested singly in 3 runs. The ratio of the mean concentration
measured over the expected concentration is expressed as a mean recovery percentage.
Sample Dilution factor Expected Mean concentration Mean recovery
concentration measured percentage (%)
(µIU/ml) (µIU/ml)
1 1/1 39.83 39.83 100
1/2 19.92 19.89 100
1/4 9.96 10.04 101
1/8 4.98 5.16 104
1/16 2.49 2.53 102
2 1/1 53.48 53.48 100
1/2 26.74 26.19 98
1/4 13.37 12.95 97
1/8 6.69 6.81 102
1/16 3.34 3.45 103
3 1/1 90.39 90.39 100
1/2 45.20 44.14 98
1/4 22.60 22.84 101
1/8 11.30 11.18 99
1/16 5.65 5.75 102

Comparison with the VIDAS TSH test method

Correlation was established between the VIDAS TSH3 kit and the VIDAS TSH kit (ref. 30 400).
The following results were obtained:
VIDAS TSH3 = 0.978 VIDAS TSH + 0.087 (n = 197)
r = 0.998
The TSH concentration of a sample determined using kits from different manufacturers, may vary depending on the test
methods used, their calibration and reagent specificity.

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WASTE DISPOSAL INDEX OF SYMBOLS

Dispose of used or unused reagents as well as any Symbol Meaning
other contaminated disposable materials following
procedures for infectious or potentially infectious Catalog number
products.
It is the responsibility of each laboratory to handle In Vitro Diagnostic Medical Device
waste and effluents produced according to their nature
and degree of hazardousness and to treat and dispose Manufacturer
of them (or have them treated and disposed of) in
accordance with any applicable regulations.
LITERATURE REFERENCES Temperature limit
1. GREEN E.D., BAENZIGER J.U. Asparagine-linked
oligosaccharides on Lutropin, Follitropin and Thyrotropin. J
Biol Chem, 1988, 263, 25-35. Use by date
2. SCANLON M.F., TOFT A.D. Regulation of Thyrotropin
Secretion. In BRAVERMAN L.E. and UTIGER R.D. eds.
th
Werner and Ingbar's The Thyroid, 7 ed. Philadelphia:
Batch code
Lippincott-Raven, 1996, 220-240.
3. SPENCER C. A., TAKEUCHI M. and KAZAROSYAN M. Consult Instructions for Use
Current status and performance goals for serum
thyrotropin (TSH) assays. Clinical Chemistry, 1996, 42 (1), Contains sufficient for <n> tests
140-145.
4. WONDISFORD F.E., MAGNER J.A. and WEINTRAUB
B.D., Chemistry and Biosynthesis of Thyrotropin. In
BRAVERMAN L.E. and UTIGER R.D. eds. Werner and Date of manufacture
th
Ingbar's The Thyroid, 7 ed. Philadelphia: Lippincott-
Raven, 1996, 190-207.
5. ZWEIG M.H., CAMPBELL G. Receiver-Operating
WARRANTY
Characteristic (ROC) plots: A Fundamental Evaluation bioMérieux disclaims all warranties, express or implied,
Tool in Clinical Medicine. Clin Chem , 1993, 39(4), 561- including any implied warranties of
577. MERCHANTABILITY AND FITNESS FOR A
PARTICULAR USE. bioMérieux shall not be liable for
any incidental or consequential damages. IN NO
EVENT SHALL BIOMERIEUX’S LIABLITY TO
CUSTOMER UNDER ANY CLAIM EXCEED A
REFUND OF THE AMOUNT PAID TO BIOMERIEUX
FOR THE PRODUCT OR SERVICE WHICH IS THE
SUBJECT OF THE CLAIM.
REVISION HISTORY
Change type categories :
N/A Not applicable (First publication)
Correction Correction of documentation anomalies
Technical change Addition, revision and/or removal of information related to the product
Administrative Implementation of non-technical changes noticeable to the user
Note: Minor typographical, grammar, and formatting changes are not included in the
revision history.
Release
Part Number Change Type Change Summary
date
INDEX OF SYMBOLS
Administrative REVISION HISTORY
2015/01 09083F CONTENT OF THE KIT (60 TESTS) – RECONSTITUTION
Technical OF REAGENTS
WARNINGS AND PRECAUTIONS
CONTENT OF THE KIT (60 TESTS) – RECONSTITUTION
2015/06 09083G Technical OF REAGENTS
INSTRUCTIONS FOR USE

BIOMERIEUX, the blue logo, SPR and VIDAS are used, pending, and/or registered trademarks belonging to bioMérieux or one of its
subsidiaries or one of its companies.
Any other name or trademark is the property of its respective owner.

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