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VIDAS® T3 (T3)
VIDAS T3 is an automated quantitative test for use on the VIDAS family instruments for the quantitative measurement of
total triiodothyronine (T3) in human serum or plasma (lithium heparin) using the ELFA technique (Enzyme Linked
Fluorescent Assay).

SUMMARY AND EXPLANATION All of the assay steps are performed automatically by the
Triiodothyronine (T3) is a hormone produced by thyroidal instrument. The reaction medium is cycled in and out of
secretion (20%) and from the peripheral deiodination the SPR several times.
mechanism which converts T4 to T3 (80%). As T3 is The sample is taken and transferred into the well
physiologically much more active than T4, it plays an containing the T3 antigen labeled with alkaline
important part in maintaining euthyroidism (1, 2). phosphatase (conjugate).
T3 circulates as a free hormone (0.3%) or bound to carrier Competition occurs between the antigen present in the
proteins (> 99.7%) such as TBG (thyroxine binding sample and the labeled antigen for the specific anti-T3
globulin), albumin or prealbumin. The free form is the antibodies (sheep) coated on the interior of the SPR.
physiologically active fraction which appears to have the Unbound components are eliminated during washing
most effect on metabolism control (3,4). steps.
T3 determination must be associated with other tests such During the final detection step, the substrate (4-Methyl-
as TSH and T4 assay, as well as with the clinical umbelliferyl phosphate) is cycled in and out of the SPR.
examination of the patient (5). The conjugate enzyme catalyzes the hydrolysis of this
substrate into a fluorescent product (4-Methyl-
PRINCIPLE umbelliferone) the fluorescence of which is measured at
450 nm. The intensity of the fluorescence is inversely
The assay principle combines an enzyme immunoassay proportional to the concentration of antigen present in the
competition method with a final fluorescent detection sample. At the end of the assay, the results are
(ELFA). automatically calculated by the instrument in relation to
®
The Solid Phase Receptacle (SPR ) serves as the solid the calibration curve stored in memory, and then printed
phase as well as the pipetting device for the assay. out.
Reagents for the assay are ready-to-use and pre-
dispensed in the sealed reagent strips.

CONTENT OF THE KIT (60 TESTS):

60 T3 strips STR Ready-to-use.
60 T3 SPRs SPR Ready-to-use.
2 x 30 Interior of SPRs coated with anti-T3 monoclonal antibodies (sheep).
T3 control C1 Ready-to-use.
1 x 2 ml (liquid) Human serum* + L-triiodothyronine + 1 g/l sodium azide.
MLE data indicate the confidence interval in nmol/L (“Control C1 Dose
Value Range”).
T3 calibrator S1 Ready-to-use.
1 x 2 ml (liquid) Human serum* + L-triiodothyronine + 1 g/l sodium azide.
MLE data indicate the concentration in nmol/L (“Calibrator (S1) Dose
Value”) and the confidence interval in "Relative Fluorescence Value”
(“Calibrator (S1) RFV Range”).
Specifications for the factory master data required to calibrate the test:
• MLE data (Master Lot Entry) provided in the kit,
or
• MLE bar codes 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, the usual
safety procedures should be observed when handling.

The SPR The strip

The interior of the SPR is coated during production with The strip consists of 10 wells covered with a labeled, foil
anti-T3 monoclonal antibodies (sheep). Each SPR is seal. The label comprises a bar code which mainly
identified by the T3 code. Only remove the required indicates the assay code, kit lot number and expiration
number of SPRs from the pouch and carefully reseal the date. The foil of the first well is perforated to facilitate the
pouch after 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.

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Description of the T3 strip

Wells Reagents
1 Sample well.
2-3-4-5 Empty wells.
6 Conjugate: alkaline phosphatase labeled T3 derivative + ANS (0.95 mmol/l) +
sodium salicylate (11.9 mmol/l) + 1 g/l sodium azide (400 µl).
7-8-9 Wash buffer: Tris-Tween-NaCl (1 mmol/l) pH 7.4 + 1 g/l of sodium azide (600 µl).
10 Cuvette with substrate: 4-methyl-ombelliferyl phosphate (0.6 mmol/l) +
diethanolamine (DEA*) (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.

MATERIALS AND DISPOSABLES REQUIRED BUT • The substrate in well 10 contains an irritant agent
NOT PROVIDED (diethanolamine). Refer to the hazard statements "H"
- Pipette with disposable tip to dispense 100 µl. and the precautionary statements "P" above.
- Powderless, disposable gloves. • Spills should be wiped up thoroughly after treatment
- For other specific materials and disposables, please with liquid detergent or a solution of household bleach
refer to the Instrument User’s Manual. containing at least 0.5% sodium hypochlorite. See the
- Instrument of the VIDAS family. User's Manual for cleaning spills on or in the instrument.
Do not autoclave solutions containing bleach.
WARNINGS AND PRECAUTIONS • The instrument should be regularly cleaned and
• For in vitro diagnostic use only. decontaminated (see the User's Manual).
• For professional use only.
• This kit contains products of human origin. No STORAGE CONDITIONS
known analysis method can totally guarantee the • Store the VIDAS T3 kit at 2-8°C.
absence of transmissible pathogenic agents. It is • Do not freeze reagents.
therefore recommended that these products be • Store all unused reagents at 2-8°C.
treated as potentially infectious, and handled • After opening the kit, check that the SPR pouch is
observing the usual safety precautions (see correctly sealed and undamaged. If not, do not use the
Laboratory Biosafety Manual - WHO - Geneva - SPRs.
latest Edition). • Carefully reseal the pouch with the desiccant inside
• This kit contains products of animal origin. Certified after use to maintain the stability of the SPRs and
knowledge of the origin and/or sanitary state of the return the complete kit to 2-8°C.
animals does not totally guarantee the absence of • If stored according to the recommended conditions, all
transmissible pathogenic agents. It is therefore components are stable until the expiration date indicated
recommended that these products be treated as on the label.
potentially infectious and handled observing the usual
safety precautions (do not ingest or inhale).
SPECIMENS
• Do not use SPRs if the pouch is pierced.
• Do not use visibly deteriorated STRs (damaged foil or Specimen type and collection:
plastic). Serum or plasma (lithium heparin). Do not use tubes
• Do not use reagents after the expiration date indicated with EDTA.
on the label.
• Do not mix reagents (or disposables) from different lots. None of the following factors have been found to
• Use powderless gloves as powder has been reported significantly influence this assay.
to cause false results for certain enzyme immunoassay - hemolysis (after spiking samples with hemoglobin: from
tests. 0 to 300 µmol/l (monomer)),
• Kit reagents contain sodium azide which can react with - lipemia (after spiking samples with lipids: from 0 to 2 g/l
lead or copper plumbing to form explosive metal azides. equivalent in triglycerides),
If any liquid containing sodium azide is disposed of in - bilirubinemia (after spiking samples with bilirubin: from 0
the plumbing system, drains should be flushed with to 320 µmol/l).
water to avoid build-up.

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However, it is recommended not to use samples that are 9. Reclose the vials and return them to
clearly hemolyzed, lipemic or icteric and, if possible, to 2–8°C after pipetting.
collect a new sample. 10. The assay will be completed within approximately 40
Specimen stability minutes. After the assay is completed, remove the
SPRs and strips from the instrument.
Samples can be stored at 2-8°C in stoppered tubes for up
to 48 hours. If longer storage is required, freeze the sera 11. Dispose of the used SPRs and strips into an
or plasma at - 25 ± 6°C. A study performed on frozen appropriate recipient.
samples over a period of 2 months, showed that the
quality of results is not affected. RESULTS AND INTERPRETATION
Avoid successive freezing and thawing. Once the assay is completed, results are analyzed
automatically by the computer. Fluorescence is measured
INSTRUCTIONS FOR USE twice in the Reagent Strip’s reading cuvette for each
For complete instructions, see the User’s Manual. sample tested. The first reading is a background reading
of the substrate cuvette before the SPR is introduced into
Reading Master lot data
the substrate. The second reading is taken after
Before each new lot of reagents is used, enter the incubating the substrate with the enzyme remaining on
specifications (or factory master data) into the instrument the interior of the SPR. The RFV (Relative Fluorescence
using the master lot entry (MLE) data. Value) is calculated by subtracting the background
If this operation is not performed before initiating the reading from the final result. This calculation appears on
tests, the instrument will not be able to print results. the result sheet.
Results are calculated automatically by the instrument in
Note: the master lot data need only be entered once relation to the calibration curve stored in memory
for each lot. (4-parameter logistic model) and are expressed in nmol/l.
It is possible to enter MLE data manually or Samples with a concentration greater than 9 nmol/l, may
automatically depending on the instrument (refer to the be diluted by 1/2 in C1 control or normal serum. The result
User’s Manual). will be calculated taking into account the dilution factor
and the concentration of C1 or normal serum used.
Calibration The results of a VIDAS T3 assay must be interpreted as
Calibration, using the calibrator provided in the kit, must part of a complete clinical profile and in association with
be performed each time a new lot of reagents is opened, thyroid function tests, including at least a TSH assay.
after the master lot data have been entered. Calibration
should then be performed every 14 days. This operation QUALITY CONTROL
provides instrument-specific calibration curves and A control is included in each VIDAS T3 kit.
compensates for possible minor variations in assay signal This control must be performed immediately after opening
throughout the shelf-life of the kit. a new kit to ensure that reagent performance has not
The calibrator, identified by "S1", must be tested in been altered. Each calibration must also be checked
triplicate (see User's Manual). The calibration value must using this control. The instrument will only be able to
be within the set RFV (Relative Fluorescence Value). If check the control value if it is identified by C1.
this is not the case: recalibrate. Results cannot be validated if the control value deviates
from the expected values.
Test procedure
Note
1. Only remove the required reagents from the
refrigerator and allow them to come to room It is the responsibility of the user to perform Quality
temperature for at least 30 minutes. Control in accordance with any local applicable
regulations.
2. Use one "T3" strip and one "T3" SPR for each sample,
control or calibrator to be tested. Make sure the
LIMITATIONS OF THE METHOD
storage pouch has been carefully resealed after
the required SPRs have been removed. Interference may be encountered with certain sera
containing antibodies directed against reagent
3. The test is identified by the "T3" code on the
components. For this reason, assay results should be
instrument. The calibrator must be identified by "S1",
interpreted taking into consideration the patient's history,
and tested in triplicate. If the control is to be tested, it
and the results of any other tests performed.
should be identified by "C1".
4. If necessary, clarify samples by centrifugation. EXPECTED VALUES
5. Mix the calibrator, control and samples using a vortex These figures are given as a guide. It is recommended
type mixer (for serum or plasma separated from the that each laboratory establishes its own reference values
pellet). from a rigorously selected population.
6. For this test, the calibrator, control, and sample As a guideline, 95% of values corresponding to 148 adults
test portion is 100 µl. who are clinically or biologically euthyroid without any
7. Insert the "T3" SPRs and "T3" strips into the associated serious illness are within the range:
instrument. Check to make sure the color labels with 0.92 - 2.33 nmol/l.
the assay code on the SPRs and the Reagent Strips
match. PERFORMANCE
8. Initiate the assay as directed in the User's Manual. All Studies performed using the VIDAS T3 assay gave the
the assay steps are performed automatically by the following results:
instrument.

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Measurement range Accuracy

The measurement range of the VIDAS T3 is: 0.4-9 nmol/l.
Dilution test:
Analytical detection limit
Three samples were diluted in human serum containing
Defined as the smallest concentration of T3 which is normal T3 concentrations and tested singly in 3 runs. The
significantly different from the zero concentration with a ratio of the mean concentration measured over the
probability of 95% < 0.4 nmol/l. expected concentration is expressed as a mean recovery
percentage.
Precision
Sample Dilution Expected Measured Recovery
Within-run reproducibility:
factor concentration concentration percentage
(nmol/l) (nmol/l)
Five samples were tested 30 times in the same run.
1 1:1 6.33 6.33 100
Sample 1 2 3 4 5 1:2 4.07 4.44 109
0.68 1.80 3.16 6.17 8.71 1:4 2.93 3.13 107
Mean concentration
1:8 2.37 2.57 109
(nmol/l)
2 1:1 7.53 7.53 100
CV % 11.0 5.3 4.4 2.7 2.3 1:2 4.53 5.22 115
1/4 3,02 3,63 120
Between-run reproducibility 1/8 2,27 2,55 112
3 1:1 3.99 3.99 100
Five samples were tested singly in 29 different runs over a 1:2 2,76 2,90 105
period of 8 weeks on the same VIDAS. 1:4 2.14 2.15 101
Sample 1 2 3 4 5 1:8 1.83 1.83 100

Mean concentration 0.76 1.93 3.34 5.84 6.42

Comparison with other test methods
(nmol/l)
199 samples (heparined serum and plasma) were
CV % 12.4 5.7 4.2 2.8 3.2 assayed in parallel using VIDAS T3 (Y) and a radio-
immunoassay kit (X).
Specificity The allometric curve equation obtained is:
The specificity of anti-T3 antibody used in this kit is: Y = 1.01 X + 0.35
Tested compound Cross-reactivity (%) r = 0.95 (n = 199)

L-Triiodothyronine 100 WASTE DISPOSAL

D-Triiodothyronine 100 Dispose of used or unused reagents as well as any other
L-Thyroxine 0.21 contaminated disposable materials following procedures
for infectious or potentially infectious products.
D-Thyroxine 0.04 It is the responsibility of each laboratory to handle waste
Diido-L-thyronine 3.3 and effluents produced according to their nature and
degree of hazardousness and to treat and dispose of
Monoiodotyrosine < 0.01 them (or have them treated and disposed of) in
Diiodotyrosine < 0.01 accordance with any applicable regulations.
Diphenylhydantoin < 0.01
LITERATURE REFERENCES
Propylthiouracile < 0.01
1. BECKER C. Thyroid hormone synthesis and circulating
Triiodothyroacetic acid 100 thyroid hormones. In thyroid diseases, World federation of
nuclear medicine and biology. Ed; C. Beckers, Pergamon
Triiodothyropropionic acid Press, 1982, 1-21.
100
2. RACADOT A. Biosynthèse des hormones thyroïdiennes.
Sodium salicylate < 0.01 Aspects biochimiques. Immunoanal. Biol. Spéc., 1991, 30,
27-32.
Phenyl butazone < 0.01
3. BIERSACK H.J. and HOTZE A. The clinician and the thyroid.
Propionic acid 50 Eur. J. Nucl. Med., 1991, 18, 761-778.
4. EVERED D.C., TUNBRIDGE W.M.G., HALL R.,
APPLETON D., BREWIS M., CLARK F., MANUEL P.,
YOUNG E. Thyroid hormone concentrations in a large scale
community survey. Effect of age, sex, illness and medication.
Clinica Chimica Acta, 1978, 223-229.
5. CARAYON P., NICCOLI-SIRE P., LEJEUNE P.J., et al. -
Recommandation de consensus sur le diagnostic et la
surveillance des maladies de la glande thyroïde. Ann. Biol.
Clin.- mai-juin 2002, vol. 60, n°3.

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INDEX OF SYMBOLS WARRANTY

Symbol Meaning bioMérieux disclaims all warranties, express or implied,
including any implied warranties of MERCHANTABILITY
Catalog number AND FITNESS FOR A PARTICULAR USE. bioMérieux
shall not be liable for any incidental or consequential
In Vitro Diagnostic Medical Device damages. IN NO EVENT SHALL BIOMERIEUX’S
LIABLITY TO CUSTOMER UNDER ANY CLAIM
Manufacturer EXCEED A REFUND OF THE AMOUNT PAID TO
BIOMERIEUX FOR THE PRODUCT OR SERVICE
WHICH IS THE SUBJECT OF THE CLAIM.
Temperature limit

Use by date

Batch code

Consult Instructions for Use

Contains sufficient for <n> tests

Date of manufacture

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
Administrative INDEX OF SYMBOLS
REVISION HISTORY
2015/01 07204I CONTENT OF THE KIT (60 TESTS)
Technical WARNINGS AND PRECAUTIONS
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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