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

SUMMARY AND EXPLANATION PRINCIPLE

LH (luteinizing hormone or luteotropin) is a glycoprotein The assay principle combines an enzyme immunoassay
with a molecular weight of approximately 30,000 daltons. sandwich method with a final fluorescent detection
Two polypeptide subunits (alpha and beta), bound by non- (ELFA).
®
covalent bonds, form LH. The LH alpha subunit, The Solid Phase Receptacle (SPR ) serves as the solid
composed of 92 amino acids, contains an amino acid phase as well as the pipetting device for the assay.
sequence similar to that of FSH, TSH and hCG. The beta Reagents for the assay are ready-to-use and pre-
subunit, composed of 121 amino acids, contains a unique dispensed in the sealed reagent strips.
amino acid sequence which gives biological and All of the assay steps are performed automatically by the
immunological specificity to LH. The LH molecule also instrument. The reaction medium is cycled in and out of
comprises carbohydrates, which account for 20% of the the SPR several times.
molecular weight of the hormone (1, 2). The sample is taken and transferred into the well
LH is secreted in a pulsatile manner by gonadotropic cells containing alkaline phosphatase-labeled anti-LH
of the anterior pituitary, controlled by the hypothalamus. antibodies (conjugate). The sample/conjugate mixture is
LH stimulates the production of steroid hormones cycled in and out of the SPR several times to increase the
(testosterone in men, and estradiol in women), which in reaction speed. The antigen binds to the antibodies
turn have a negative feedback effect on gonadotropins. In coated on the SPR and to the conjugate forming a
males, LH stimulates the production of testosterone by the "sandwich".
Leydig cells. In menstruating women, LH, in synergy with Unbound components are eliminated during the washing
FSH, directly affects the development of ovarian follicles, steps. During the final detection step, the substrate
increasing their steroid production and determining (4-Methyl-umbelliferyl phosphate) is cycled in and out of
ovulation. At menopause, estradiol production falls the SPR. The conjugate enzyme catalyzes the hydrolysis
drastically, resulting in a rise in the serum level of LH of this substrate into a fluorescent product
(3, 4). (4-Methyl-umbelliferone) the fluorescence of which is
The LH assay, whether or not combined with FSH, is an measured at 450 nm. The intensity of the fluorescence is
essential parameter for the investigation of the proportional to the concentration of antigen present in the
reproductive function. For men and amenorrheic women, sample.
elevated LH levels indicate primary hypogonadism while At the end of the assay, the results are automatically
low LH levels indicate secondary hypogonadism (5, 6, 7). calculated by the instrument in relation to the calibration
curve stored in memory, and then printed out.
CONTENT OF THE KIT (60 TESTS) - RECONSTITUTION OF REAGENTS:
60 LH strips STR Ready-to-use.
60 LH SPRs SPR Ready-to-use.
2 x 30 Interior of SPRs coated with monoclonal anti-LH immunoglobulins (mouse).
LH control C1 Reconstitute with 3 ml of distilled water. Leave to stand for 5 to 10 minutes, then mix.
1 x 3 ml Stable after reconstitution for 14 days at 2-8°C or until expiration date on the kit at -
(lyophilized) 25  6°C.
Five freeze/thaw cycles are possible.
Human serum* + human LH + preservatives.
MLE data indicate the confidence interval in mIU/mL (milli-international units per milliliter)
(“Control C1 Dose Value Range”).
LH calibrator S1 Reconstitute with 2 ml of distilled water. Leave to stand for 5 to 10 minutes, then mix.
3 x 2 ml Stable after reconstitution for 14 days at 2-8°C or until expiration date on the kit at -
(lyophilized) 25  6°C.
Five freeze/thaw cycles are possible.
Phosphate buffer (0.05 mol/l, pH 7.5) + human LH + protein and chemical stabilizers +
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”).
LH dilution buffer R1 Ready-to-use.
1 x 3 ml (liquid) Phosphate buffer (0.05 mol/l, pH 7.5) + protein and chemical stabilizers + 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 [Link]/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, the usual
safety procedures should be observed when handling.

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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-LH immunoglobulins. Each SPR seal. The label comprises a bar code which mainly
is identified by the LH 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 LH reagent strip
Wells Reagents
1 Sample well.
2-3-4-5 Empty wells.
6 Conjugate: alkaline phosphatase-labeled monoclonal anti-LH 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 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

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 WARNINGS AND PRECAUTIONS
NOT PROVIDED  For in vitro diagnostic use only.
- Pipette with disposable tip to dispense 3ml, 2ml and  For professional use only.
200 µl.
- Powderless, disposable gloves.
- For other specific materials and disposables, please
refer to the Instrument User’s Manual.
- Instrument of the VIDAS family.

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 This kit contains products of human origin. No - bilirubinemia (after spiking samples with bilirubin:
known analysis method can totally guarantee the 0 to 513 µmol/l).
absence of transmissible pathogenic agents. It is However, it is recommended not to use samples that are
therefore recommended that these products be clearly hemolyzed, lipemic or icteric and, if possible, to
treated as potentially infectious and handled collect a new sample.
observing the usual safety precautions (see
Laboratory Biosafety Manual - WHO - Geneva - Specimen stability
latest Edition). Samples can be stored at 2-8°C in stoppered tubes for up
 This kit contains products of animal origin. Certified to 48 hours; if longer storage is required, freeze the sera
knowledge of the origin and/or sanitary state of the or plasma at -25 ° 6 C.
animals does not totally guarantee the absence of Avoid successive freezing and thawing.
transmissible pathogenic agents. It is therefore
recommended that these products be treated as INSTRUCTIONS FOR USE
potentially infectious and handled observing the usual For complete instructions, see the User’s Manual.
safety precautions (do not ingest or inhale).
Reading Master lot data
 Do not use the SPRs if the pouch is pierced.
 Do not use visibly deteriorated STRs (damaged foil or Before each new lot of reagents is used, enter the
plastic). specifications (or factory master data) into the instrument
 Do not use reagents after the expiration date indicated using the master lot entry (MLE) data.
on the label. If this operation is not performed before initiating the
 Do not mix reagents (or disposables) from different lots. tests, the instrument will not be able to print results.
 Use powderless gloves, as powder has been reported
to cause false results for certain enzyme immunoassay Note: the master lot data need only be entered once
tests. for each lot.
 Kit reagents contain sodium azide which can react with It is possible to enter MLE data manually or
lead or copper plumbing to form explosive metal azides. automatically depending on the instrument (refer to the
If any liquid containing sodium azide is disposed of in User’s Manual).
the plumbing system, drains should be flushed with
water to avoid build-up. Calibration
 The wash buffer (well 9) contains a harmful agent Calibration, using the calibrator provided in the kit, must
(11.5% diethanolamine). Refer to the hazard statements be performed each time a new lot of reagents is opened,
"H" and the precautionary statements "P" above. after the master lot data have been entered. Calibration
 The substrate (well 10) contains an irritant agent should then be performed every 14 days. This operation
(6.6% diethanolamine). Refer to the hazard statements provides instrument-specific calibration curves and
"H" and the precautionary statements "P" above. compensates for possible minor variations in assay signal
 Spills should be wiped up thoroughly after treatment throughout the shelf-life of the kit.
with liquid detergent or a solution of household bleach The calibrator, identified by S1, must be tested in
containing at least 0.5% sodium hypochlorite. See the duplicate (see User’s Manual). The calibrator value must
User’s Manual for cleaning spills on or in the instrument. be within the set RFV "Relative Fluorescence Value"
Do not autoclave solutions containing bleach. range. If this is not the case, recalibrate.
 The instrument should be regularly cleaned and
Procedure
decontaminated (see the User’s Manual).
STORAGE CONDITIONS 1. Only remove the required reagents from the
refrigerator and allow them to come to room
 Store the LH VIDAS kit at 2-8°C.
temperature for at least 30 minutes.
 Do not freeze SPRs, reagent strips or dilution buffer.
 Store all unused reagents at 2-8°C. 2. Use one "LH" strip and one "LH" SPR for each
 After opening the kit, check that the SPR pouch is sample, control or calibrator to be tested. Make sure
correctly sealed and undamaged. If not, do not use the that the storage pouch has been carefully resealed
SPRs. after the required SPRs have been removed.
 Carefully reseal the pouch with the desiccant inside 3. The test is identified by the "LH" code on the
after use to maintain stability of the SPRs and return instrument. The calibrator must be identified by "S1",
the complete kit to 2-8°C. and tested in duplicate. If the control needs to be
 If stored according to the recommended conditions, all tested, it should be identified by "C1".
components are stable until the expiration date indicated 4. If necessary, clarify samples by centrifugation.
on the label. Refer to the kit composition table for
special storage conditions. 5. Mix the calibrator, control and samples using a vortex-
type mixer (for serum or plasma separated from the
SPECIMENS pellet).
Specimen type and collection 6. For this test, the calibrator, control, and sample
Serum or plasma (lithium heparin). Do not use plasma test portion is 200 µl.
collected on EDTA.
7. Insert the "LH" SPRs and "LH" strips into the
None of the following factors have been found to instrument. Check to make sure the color labels with
significantly influence this assay. the assay code on the SPRs and the Reagent strips
- hemolysis (after spiking samples with hemoglobin: match.
0 to 300 µmol/l (monomer)),
8. Initiate the assay as directed in the User’s Manual. All
- lipemia (after spiking samples with lipids: 0 to 2 g/l
the assay steps are performed automatically by the
equivalent in triglycerides),
instrument.

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9. Reclose the vials and return them to the required RANGE OF EXPECTED VALUES
temperature after pipetting. These figures are given as a guide; it is recommended
10. The assay will be completed within approximately that each laboratory establishes its own reference values
40 minutes. After the assay is completed, remove the from a rigorously selected population.
SPRs and strips from the instrument. Men: 1.1 - 7.0 mIU/ml
11. Dispose of used SPRs and reagent strips in an Women:
appropriate recipient.
- Ovulation peak (D0) 9.6 - 80.0 mIU/ml
RESULTATS AND INTERPRETATION - Follicular phase:
Once the assay is completed, results are analyzed First half (D-15 to D-9) 1.5 - 8.0 mIU/ml
automatically by the computer. Fluorescence is measured Second half (D-8 to D-2) 2.0 - 8.0 mIU/ml
twice in the Reagent Strip’s reading cuvette for each - Luteal phase (D+3 to D+15) 0.2 - 6.5 mIU/ml
sample tested. The first reading is a background reading Menopause: 8.0 - 33.0 mIU/ml
of the substrate cuvette before the SPR is introduced into
the substrate. The second reading is taken after PERFORMANCE
incubating the substrate with the enzyme remaining on Studies performed using VIDAS LH gave the following
the interior of the SPR. The RFV (Relative Fluorescence results:
Value) is calculated by subtracting the background
reading from the final result. This calculation appears on Measurement range
the result sheet. The measurement range of the VIDAS LH reagent
The results are automatically calculated by the instrument extends from 0.1-100 mIU/ml.
using calibration curves which are stored by the
instrument (4-parameter logistics model). The LH Detection limit
concentrations are expressed in mIU/ml (2nd IS 80/552). Defined as the smallest concentration of LH which is
Samples with a LH concentration greater than 100 mIU/ml significantly different from the zero concentration with a
should be retested after dilution to 1/2 or 1/4 in LH dilution probability of 95%: < 0.1 mIU/ml.
buffer (R1). Multiply the result by the dilution factor to
obtain the sample concentration. Hook effect
Interpretation of test results should be made taking into No hook effect was found up to LH concentrations of
consideration the patient’s history, and any other tests 10,000 mIU/ml.
performed.
Precision
QUALITY CONTROL
Within-run reproducibility:
A control is included in each LH VIDAS kit.
This control must be performed immediately after opening 5 samples were tested 30 times in a same run.
a new kit to ensure that reagent performance has not Sample 1 2 3 4 5
been altered. Each calibration must also be checked
using this control. The instrument will only be able to Mean concentration
1.6 7.1 25.9 37.8 66.1
check the control value if it is identified by C1. (mIU/ml)
Results cannot be validated if the control value deviates CV% 4.5 4.8 3.8 4.3 3.1
from the expected values.
Between-run reproducibility:
Note
5 samples were tested singly in 24 different runs on the
It is the responsibility of the user to perform Quality
same VIDAS over a period of 8 weeks.
Control in accordance with any local applicable regulation.
Sample 1 2 3 4 5
LIMITATIONS OF THE METHOD Mean concentration
1.7 7.2 26.3 33.3 64.7
Interference may be encountered with certain sera (mIU/ml)
containing antibodies directed against reagent
CV % 3.8 6.6 4.0 4.5 3.7
components. For this reason, assay results should be
interpreted taking into consideration the patient history,
and the results of any other tests performed. Specificity
Tested compound Cross-reactions (%)
LH 100
FSH < 0.1
TSH < 0.1
hCG < 0.01
hCG  subunit < 0.01

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Accuracy
Dilution test
Three samples were diluted in the LH dilution buffer (R1) and tested singly in 3 series. The ratio of the mean
concentration measured over the expected concentration is expressed as a mean recovery percentage.
Expected Mean measured Mean recovery
Sample no. Dilution factor concentration concentration percentage
(mIU/ml) (mIU/ml) (%)
1/1 30.3 30.33 100
1/2 15.2 15.19 100
1 1/4 7.6 7.27 96
1/8 3.8 3.51 92
1/16 1.9 1.76 93
1/32 0.9 0.84 93
1/1 39.9 39.91 100
1/2 20.0 20.92 105
2 1/4 10.0 9.76 98
1/8 5.0 5.14 103
1/16 2.5 2.53 101
1/32 1.2 1.30 108
1/1 43.9 43.95 100
1/2 22.0 24.50 111
3 1/4 11.0 11.57 105
1/8 5.5 5.84 106
1/16 2.7 3.06 113
1/32 1.4 1.44 103

Comparison with other test methods LITERATURE REFERENCES

135 serum samples were tested using VIDAS (Y) and a 1. BARDIN C.W. et PAULSEN C.A. Textbook of endocrinology
radioimmunoassay kit (X). - Ed WILLIAMS H.R. et SAUNDERS W.B., 6ème édition
The allometric curve equation obtained is: Philadelphia, 1981 - The Testes, 6, 293-354.
Y = 0.929 X - 0.53 with a correlation coefficient of 0.987. 2. BUTT W.R. Hormones in blood - Ed GRAY C.H. et JAMES
W.T.H., 3ème édition, Academic Press, London, 1983,
WASTE DISPOSAL Gonadotrophins - 7, 147-177.
Dispose of used or unused reagents as well as any other 3. LANDGREN B.M., UNDEN A.L. et DICZFALUSY E.
contaminated disposable materials following procedures Hormonal profile of the cycle in 68 normal menstruating
for infectious or potentially infectious products. women. Acta Endocrinologica, 1980, 94, 89-98.
It is the responsibility of each laboratory to handle waste 4. ROSS G.T., VANDE WIELE R.L. et FRANTZ A.G. -
and effluents produced according to their nature and Textbook of endocrinology - Ed WILLIAMS R. H. et
SAUNDERS W.B. Philadelphia 1981 - The Ovaries and the
degree of hazardousness and to treat and dispose of Breasts - 6, 355-411.
them (or have them treated and disposed of) in
5. SCHOLLER R., Ed. Hormonologie de la Stérilité. Exploration
accordance with any applicable regulations.
et thérapeutique. Journées d'endocrinologie clinique du 21-
22 novembre 1980, Edition SEPE Paris – Taux des gonado-
tropines et des stéroïdes chez la femme normale - 165-193.
6. TESTART J., FRYDMAN R., BELAISCH J.C., CASTANIER
M., FEINSTEIN M.C., GRENIER J. et ROGER M. Profil
hormonal plasmatique et intrafolliculaire au cours de la
période préovulatoire tardive. J. Gyn. Obst. Biol. Repr., 1983,
12, 119-127.
7. WIDE L. Hormone assays and their clinical applications - Ed
LORAINE J.A. et BELL E.T., 4ème édition, Churchill
Livingstone, Edinburgh, London and New York, 1976 -
Human Pituitary Gonadotrophins - 87-140.

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INDEX OF SYMBOLS WARRANTY

Symbol Meaning bioMérieux disclaims all warranties, express or implied,
including any implied warranties of MERCHANTABILITY
AND FITNESS FOR A PARTICULAR USE. bioMérieux
Catalog number
shall not be liable for any incidental or consequential
damages. IN NO EVENT SHALL BIOMERIEUX’S
In Vitro Diagnostic Medical Device LIABILITY TO CUSTOMER UNDER ANY CLAIM
EXCEED A REFUND OF THE AMOUNT PAID TO
BIOMERIEUX FOR THE PRODUCT OR SERVICE
Manufacturer 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
INDEX OF SYMBOLS
Administrative REVISION HISTORY
2015/01 06267O CONTENT OF THE KIT (60 TESTS) – RECONSTITUTION
Technical OF REAGENTS
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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