30 407 06268 M - en - 2015/01

VIDAS®FSH (FSH)
VIDAS FSH is an automated quantitative test for use on the VIDAS family instruments for the determination of human
follicle stimulating hormone in human serum or plasma (lithium heparin), using the ELFA technique (Enzyme Linked
Fluorescent Assay).

SUMMARY AND EXPLANATION PRINCIPLE

Follicle-Stimulating Hormone (FSH) is a glycoprotein with The assay principle combines an enzyme immunoassay
a molecular weight of about 30,000 daltons. Two sandwich method with a final fluorescent detection
polypeptide subunits (alpha and beta), bound by non- (ELFA).
®
covalent bonds, form FSH. The alpha subunit, composed The Solid Phase Receptacle (SPR ), serves as the solid
of 92 amino acids, is similar to that of LH, TSH, and hCG. phase as well as the pipetting device for the assay.
The beta subunit, composed of 111 amino acids, gives Reagents for the assay are ready-to-use and pre-
biological and immunological specificity to FSH. The FSH dispensed in the sealed reagent strips.
molecule also comprises carbohydrates, which account for All of the assay steps are performed automatically by the
20 % of the molecular weight of the hormone (1, 2). instrument. The reaction medium is cycled in and out of
FSH is secreted in a pulsatile manner by gonadotropic the SPR several times.
cells of the anterior pituitary, controlled by the The sample is taken and transferred into the well
hypothalamus. FSH stimulates production of inhibin, which containing alkaline phosphatase-labeled anti-FSH
in turn controls FSH production through a negative (conjugate). The sample/conjugate mixture is cycled in
feedback mechanism. and out of the SPR several times to increase the reaction
In men, FSH acts upon the seminiferous tubules (Sertoli speed. The antigen binds to antibodies coated on the SPR
cells). Its action on the Sertoli cells stimulates the and to the conjugate forming a "sandwich".
production of Androgen Binding Protein (ABP) and inhibin. Unbound components are eliminated during the washing
In women, with normal menstrual cycles, FSH in synergy steps. During the final detection step, the substrate
with LH, stimulates the development of the ovarian (4-Methyl-umbelliferyl phosphate) is cycled in and out of the
follicles, increasing their steroid production and SPR. The conjugate enzyme catalyzes the hydrolysis of this
dertermining ovulation. Furthermore, FSH stimulates substrate into a fluorescent product (4-Methyl-umbelliferone),
certain follicular lutein cells, leading to the production of the fluorescence of which is measured at 450 nm. The
inhibin. During menopause, since there is a sharp drop in intensity of the fluorescence is proportional to the
the production of inhibin, a significant increase in serum concentration of antigen present in the sample.
FSH is observed (3, 4). At the end of the assay, results are automatically
The measurement of FSH, sometimes in conjunction with calculated by the instrument in relation to the calibration
that of LH, is a major parameter to examine the function of curve stored in memory, and then printed out.
reproduction.
In men or in amenorrheic women, high rates are a sign of
primary hypogonadism, and low rates indicate secondary
hypogonadism (5, 6).

bioMérieux SA English - 1
 ®
VIDAS FSH (FSH) 06268 M - en - 2015/01

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

60 FSH strips STR Ready-to-use.
60 FSH SPRs SPR Ready-to-use. Interior of SPRs coated with monoclonal anti-FSH immunoglobulins
2 x 30 (mouse).
FSH control C1 Reconstitute with 3 ml of distilled water. Leave to stand for 5 to 10 minutes, then mix.
1 x 3 ml (lyophilized) Stable after reconstitution for 14 days at 2-8°C or until the expiration date on the
kit at - 25 ± 6°C.
5 freeze/thaw cycles are possible.
Human serum* + human FSH + preservatives.
MLE data indicate the confidence interval in mIU/mL (milli-international units per
milliliter) ("Control C1 Dose Value Range").
FSH calibrator S1 Reconstitute with 2 ml of distilled water. Leave to stand for 5 to 10 minutes, then mix.
3 x 2 ml (lyophilized) Stable after reconstitution for 14 days at 2-8°C or until the expiration date on the
kit at - 25 ± 6°C.
5 freeze/thaw cycles are possible
Bovine serum + human FSH + preservatives.
MLE data indicate the concentration in mIU/mL ("Calibrator (S1) Dose Value") and the
confidence interval in "Relative Fluorescence Value ("Calibrator (S1) RFV Range).
FSH diluent R1 Ready-to-use.
1 x 3 ml (liquid) Bovine serum + 1 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, and 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 interior of the SPR is coated during production with The strip consists of 10 wells covered with a labeled, foil
mouse monoclonal anti-FSH immunoglobulins. Each SPR seal. The label comprises a bar code which mainly
is identified by the FSH 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.

Description of the VIDAS FSH strip

Wells Reagents
1 Sample well.
2-3-4-5 Empty wells.
6 Conjugate: alkaline phosphatase-labeled monoclonal anti-FSH immunoglobulins
(mouse) + 1 g/l sodium azide (400 µl).
7-8 Wash buffer: sodium phosphate (0.01 mol/l) pH 7.4 + 1 g/l sodium azide (600 µl).
9 Wash buffer: diethanolamine* (1.1 mol/l or 11.5 %) pH 9.8 + 1 g/l sodium azide
(600 µl).
10 Reading cuvette with substrate: 4 Methyl-umbelliferyl-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

bioMérieux SA English - 2
 ®
VIDAS FSH (FSH) 06268 M - en - 2015/01

Hazard statement
H318 : Causes serious eye damage.
H373 : May cause damage to organs through prolonged or repeated exposure.
H315 : Causes skin irritation.
H302 : Harmful if swallowed.
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.
P309 + P311 : IF exposed or if you feel unwell: Call a POISON CENTER or doctor/physician.
** 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 wash buffer (well 9) contains a harmful agent
NOT PROVIDED (11.5% diethanolamine). Refer to the hazard statements
- Pipette with disposable tip to dispense 2 ml, 3 ml and "H" and the precautionary statements "P" above.
200 µl. • The substrate (well 10) contains an irritant agent (6.6%
- Powderless, disposable gloves. diethanolamine). Refer to the hazard statements "H" and
- For other specific materials and disposables, please the precautionary statements "P" above.
refer to the Instrument User’s Manual • Spills should be wiped up thoroughly after treatment with
- VIDAS family instrument. liquid detergent and a solution of household bleach
containing at least 0.5% sodium hypochlorite. See the
User’s Manual for cleaning spills on or in the instrument.
WARNINGS AND PRECAUTIONS
Do not autoclave solutions containing bleach.
• For in vitro diagnostic use only. • The instrument should be regularly cleaned and
• For professional use only. decontaminated (see the User’s Manual).
• This kit contains products of human origin. No
known analysis method can totally guarantee the STORAGE CONDITIONS
absence of transmissible pathogenic agents. It is
therefore recommended that these products be • Store the VIDAS FSH kit at 2-8°C.
treated as potentially infectious and handled • Do not freeze SPRs, strips and diluent.
observing the usual safety precautions (see • Store all unused reagents at 2-8°C.
Laboratory biosafety manual - WHO - Geneva - latest • After opening the kit, check that the SPR pouch is
edition). correctly sealed and undamaged. If not, do not use the
• This kit contains products of animal origin. Certified SPRs.
knowledge of the origin and/or sanitary state of the
• Carefully reseal the pouch with the desiccant inside
animals does not totally guarantee the absence of
after use to maintain stability of the SPRs and return
transmissible pathogenic agents. It is therefore
the complete kit to 2-8°C.
recommended that these products be treated as
potentially infectious and handled observing the usual • If stored according to the recommended conditions, all
safety precautions (do not ingest or inhale). components are stable until the expiration date indicated
• Do not use the SPRs if the pouch is pierced. on the label. Refer to the kit composition table for
• Do not use visibly deteriorated STRs (damaged foil or special storage conditions.
plastic).
• Do not use reagents after the expiration date indicated SPECIMENS
on the label. Specimen type and collection
• Do not mix reagents (or disposables) from different lots. Serum or plasma (lithium heparin). Do not use plasma
• Use powderless gloves, as powder has been reported collected on EDTA.
to cause false results for certain enzyme immunoassay None of the following factors have been found to
tests. significantly influence this assay.
• Kit reagents contain sodium azide which can react with - hemolysis (after spiking samples with hemoglobin:
lead or copper plumbing to form explosive metal azides. 0 to 300 µmol/l (monomer)),
If any liquid containing sodium azide is disposed of in - lipemia (after spiking samples with lipids: 0 to 2 g/l
the plumbing system, drains should be flushed with equivalent in triglycerides),
water to avoid build-up.

bioMérieux SA English - 3
 ®
VIDAS FSH (FSH) 06268 M - en - 2015/01

- bilirubinemia (after spiking samples with bilirubin: 8. Initiate the assay as directed in the User’s Manual. All
0 to 513 µmol/l). the assay steps are performed automatically by the
However, it is recommended not to use samples that are instrument.
clearly hemolyzed, lipemic or icteric and, if possible, to 9. Reclose the vials and return them to the required
collect a new sample. temperature after pipetting.
Specimen stability 10. The assay will be completed within approximately 40
Samples can be stored at 2-8°C in stoppered tubes for a minutes. After the assay is completed, remove the
maximum of 48 hours; if longer storage is required, freeze SPRs and strips from the instrument.
the sera or plasma at - 25 ± 6°C. Avoid successive
freezing and thawing. 11. Dispose of the used SPRs and strips into an
appropriate recipient.
INSTRUCTIONS FOR USE
RESULTS AND INTERPRETATION
For complete instructions, see the User’s Manual.
Once the assay is completed, results are analyzed
Reading Master lot data automatically by the computer. Fluorescence is measured
twice in the Reagent Strip’s reading cuvette for each
Before each new lot of reagents is used, enter the sample tested. The first reading is a background reading
specifications (or factory master data) into the instrument of the substrate cuvette before the SPR is introduced into
using the master lot entry (MLE) data. the substrate. The second reading is taken after
If this operation is not performed before initiating the incubating the substrate with the enzyme remaining on the
tests, the instrument will not be able to print results. interior of the SPR. The RFV (Relative Fluorescence
Note: the master lot data need only be entered once Value) is calculated by subtracting the background
for each lot. reading from the final result. This calculation appears on
the result sheet.
It is possible to enter MLE data manually or
The results are automatically calculated by the instrument
automatically depending on the instrument (refer to the
using calibration curves which are stored by the
User’s Manual).
instrument (4-parameter logistics model) and are
Calibration expressed in mIU/ml (2nd IRP 78/549).
Calibration, using the calibrator provided in the kit, must Samples with FSH concentrations greater than 110 mIU/ml
be performed each time a new lot of reagents is opened, must be reassayed after dilution in the FSH diluent (R1).
after the master lot data have been entered. Calibration Multiply the result by the dilution factor to obtain the sample
should then be performed every 14 days. This operation concentration.
provides instrument-specific calibration curves and Interpretation of test results should be made taking into
compensates for possible minor variations in assay signal consideration the patient history, and any other tests
throughout the shelf-life of the kit. A control must be tested performed.
after each calibration.
The calibrator, identified by S1, must be tested in QUALITY CONTROL
duplicate (see User’s Manual). The calibration value must A control is included in each VIDAS FSH kit.
be within the set RFV (Relative Fluorescence Value). If This control must be performed immediately after opening
this is not the case, recalibrate. a new kit to ensure that reagent performance has not
been altered. Each calibration must also be checked using
Procedure
this control.
1. Only remove the required reagents from the The instrument will only be able to check the control value
refrigerator and allow them to come to room if it is identified by C1.
temperature for at least 30 minutes. Results cannot be validated if the control value deviates
2. Use one "FSH" strip and one "FSH" SPR from the kit from the expected values.
for each sample, control or calibrator to be tested. Note
Make sure the storage pouch has been carefully
It is the responsibility of the user to perform Quality
resealed after the required SPRs have been
Control in accordance with any local applicable
removed.
regulations.
3. The test is identified by the "FSH" code on the
instrument. The calibrator must by identified by "S1"
LIMITATIONS OF THE METHOD
and tested in duplicate. If the control is to be tested, it
should be identified by "C1". Interference may be encountered with certain sera
containing antibodies directed against reagent
4. If necessary, clarify samples by centrifugation.
components. For this reason, assay results should be
5. Mix the calibrator, control and samples using a vortex- interpreted taking into consideration the patient history,
type mixer (for serum or plasma separated from the and any other tests performed.
pellet).
6. For this test, the calibrator, control, and sample
test portion is 200 µl.
7. Insert the "FSH" SPRs and "FSH" strips into the
instrument. Check to make sure the color labels with
the assay code on the SPRs and the Reagent Strips
match.

bioMérieux SA English - 4
 ®
VIDAS FSH (FSH) 06268 M - en - 2015/01

RANGE OF EXPECTED VALUES Precision

These figures are given as a guide; it is recommended Within-run reproducibility
that each laboratory establishes its own reference values
Five samples were tested 30 times in a same run.
from a rigorously selected population.
Sample 1 2 3 4 5
Men: 1.7- 12.0 mIU/ml
Mean
Women: concentration 4.0 10.5 53.5 82.7 90.8
- Ovulation peak (D0) 6.3 - 24.0 mIU/ml (mIU/ml)
- Follicular phase:
st
CV % 4.9 3.4 3.6 4.3 4.7
1 half (D-15 to D-9) 3.9 - 12.0 mIU/ml
nd
2 half (D-8 to D-2) 2.9 - 9.0 mIU/ml Between-run reproducibility
- Luteal phase (D+3 to D+15) 1.5 - 7.0 mIU/ml Five samples were tested singly in 24 different runs on the
- Menopause: 17.0 - 95.0 mIU/ml same VIDAS instrument over a 9-week period.
Sample 1 2 3 4 5
PERFORMANCE
Mean
Studies performed using VIDAS FSH gave the following
concentration 4.9 10.8 55.1 86.5 101.5
results:
(mIU/ml)
Measurement range CV % 3.5 3.9 4.8 5.6 5.9
The measurement range of the VIDAS FSH reagent
extends from 0.1 to 110 mIU/ml. Specificity
Analytical detection limit Tested compound % cross-reactions
Defined as the smallest concentration of FSH which is FSH 100
significantly different from the zero concentration with a
LH < 0.15
probability of 95%: < 0.1 mIU/ml.
TSH < 0.15
Hook effect
hCG < 0.01
No hook effect was found up to FSH concentrations of
3,000 mIU/ml. HCG α - subunit < 0.01

Accuracy - Dilution test:

Three samples were diluted in the FSH diluent (R1) and tested singly in three runs. The measured mean concentration
compared to the expected mean concentration is expressed as a mean recovery percentage.
Expected mean Mean measured
Mean recovery
Sample Dilution factor concentration concentration
percentage
(mIU/ml) (mIU/ml)
1/1 52.6 52.6 100.0
1/2 26.3 25.0 95.1
1 1/4 13.1 12.1 92.0
1/8 6.6 6.3 95.3
1/16 3.3 3.2 98.3
1/32 1.6 1.7 102.1
1/1 83.1 83.1 100.0
1/2 41.5 39.6 95.3
2 1/4 20.8 20.0 96.2
1/8 10.4 9.8 94.6
1/16 5.2 5.0 96.0
1/32 2.6 2.7 103.1
1/1 94.9 95.0 100.0
1/2 47.5 45.0 94.9
3 1/4 23.7 23.6 99.5
1/8 11.9 11.1 93.4
1/16 5.9 5.7 96.4
1/32 3.0 3.1 103.7

bioMérieux SA English - 5
 ®
VIDAS FSH (FSH) 06268 M - en - 2015/01

Comparison with other test methods INDEX OF SYMBOLS

183 serum samples were tested using the VIDAS and Symbol Meaning
another commercially available kit.
The allometric line equation obtained is: Catalog number
Y = 0.986 X - 0.09 with a correlation coefficient of
0.985. In Vitro Diagnostic Medical Device
WASTE DISPOSAL
Dispose of used or unused reagents as well as any Manufacturer
other contaminated disposable materials following
procedures for infectious or potentially infectious
products. Temperature limit
It is the responsibility of each laboratory to handle waste
and effluents produced according to their nature and
degree of hazardousness and to treat and dispose of Use by date
them (or have them treated and disposed of) in
accordance with any applicable regulations. Batch code

LITERATURE REFERENCES Consult Instructions for Use

1. BARDIN C.W. et PAULSEN C.A. Textbook of
endocrinology - Ed WILLIAMS H.R. et SAUNDERS W.B., Contains sufficient for <n> tests
6ème édition Philadelphia, 1981 - The Testes, 6, 293-354.
2. BUTT W.R. Hormones in blood - Ed GRAY C.H. et JAMES
W.T.H., 3ème édition, Academic Press, London, 1983, Date of manufacture
Gonadotrophins - 7, 147-177.
3. LANDGREN B.M., UNDEN A.L. et DICZFALUSY E.
Hormonal profile of the cycle in 68 normal menstruating
women. Acta Endocrinologica, 1980, 94, 89-98. WARRANTY
4. ROSS G.T., VANDE WIELE R.L. et FRANTZ A.G. - bioMérieux disclaims all warranties, express or implied,
Textbook of endocrinology - Ed WILLIAMS R. H. et including any implied warranties of
SAUNDERS W.B. Philadelphia 1981 - The Ovaries and the
Breasts - 6, 355-411.
MERCHANTABILITY AND FITNESS FOR A
5. WIDE L. Hormone assays and their clinical applications - PARTICULAR USE. bioMérieux shall not be liable for
Ed LORAINE J.A. et BELL E.T., 4ème édition, Churchill any incidental or consequential damages. IN NO
Livingstone, Edinburgh, London and New York, 1976 - EVENT SHALL BIOMERIEUX’S LIABILITY TO
Human Pituitary Gonadotrophins, 87-140. CUSTOMER UNDER ANY CLAIM EXCEED A
6. SCHOLLER R., Ed. Hormonologie de la Stérilité. REFUND OF THE AMOUNT PAID TO BIOMERIEUX
Exploration et thérapeutique. Journées d'endocrinologie FOR THE PRODUCT OR SERVICE WHICH IS THE
clinique du 21-22 novembre 1980, Edition SEPE Paris – SUBJECT OF THE CLAIM.
Taux des gonado-tropines et des stéroïdes chez la femme
normale – 165-193.

bioMérieux SA English - 6
 ®
VIDAS FSH (FSH) 06268 M - en - 2015/01

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
CONTENT OF THE KIT (60 TESTS) - RECONSTITUTION
2015/01 06268M
OF REAGENTS
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.

673 620 399 RCS LYON

bioMérieux SA Tél. 33 (0)4 78 87 20 00
376 Chemin de l'Orme Fax 33 (0)4 78 87 20 90
69280 Marcy-l'Etoile - France www.biomerieux.com