Elecsys LH

Elecsys® 1010/2010/MODULAR ANALYTICS E170
Product Information Elecsys® LH Immunoassay
Product Information
Elecsys® LH
Luteinizing hormone
11732234-122
Elecsys® Systems 1010/2010

MODULAR ANALYTICS E170
1
Contents Page
1 Intended Use 3
2 Summary 3
2.1 Biochemistry, Clinical Indications, Methodology 3–5

2.2 Elecsys Test Principle 6–7
3 Reagents 8
3.1 Contents and Concentrations 8

Test Reagent Kit, Calibrator Set, Controls,
System Reagents
3.2 Precautions and Warnings 8
3.3 Reagent Handling 9
Test Reagent Kit, Calibrator Set, Controls
3.4 Storage and Stability 9
Test Reagent Kit, Calibrator Set, Controls
4 Specimen Collection and Preparation 10
5 Elecsys LH Testing Procedure 10
5.1 Materials Provided, Materials Required (but not provided) 10

5.2 Assay Procedure 10
5.3 Calibration
5.3.1 Standardization Traceability 11 – 12
5.3.2 Calibration Frequency 12
5.3.3 Verification of Calibration 12
5.4 Quality Control 12
5.5 Calculation 12
5.6 Limitations – Interference 13
5.7 Measuring Range (Reportable Range) 14
5.8 Dilution 14
5.9 Expected Values (Reference Range) 14 – 15
6 Specific Performance Data of the Test 16
6.1 Precision, Within-run / Total / Inter-Instrument / Inter-Module Precision 16

6.2 Analytical Sensitivity (Lower Detection Limit) 17
6.3 Analytical Specificity 17
6.4 Method Comparisons 17 – 19
7 References 20
8 Results of Multicenter Studies 21
8.1 Precision 21
8.1.1 Within-run Precisision 21
8.1.2 Between-day and Total Precision 21
8.1.3 Inter-Instrument and Inter Module Precision 22
8.1.4 Inter-Instrument Precision, LH CalSEt II, 03561097 22
8.2 Method Comparisons 23 – 29
8.3 Clinical Data – Reference Ranges, Profiles of Typical Menstrual Cycles 30 – 32
9 Short Guide to Elecsys LH

Please note: As a result of differing printing dates, it is possible that differences may occur between the information given here and that appearing in the
package
2 inserts. In such cases the data given in the insert enclosed with the kit applies. Please also note that the product information document contains more
extensive information than the package insert.
Significant additions or changes are indicated by a changebar in the margin.
1 Intended Use
Immunoassay for the in vitro quantitative determination of luteinizing hormone (LH) in human serum
and plasma.
The electrochemiluminescence immunoassay “ECLIA” is intended for use on the Roche Diagnostics
Elecsys 1010/2010/MODULAR ANALYTCS E170 (Elecsys Modul) immunoassay analyzers.
2 Summary
2.1 Biochemistry, Biochemistry

Clinical Indications, LH (luteinizing hormone), together with FSH (follicle stimulating hormone), belongs to the gonadotropin
Methodology family. LH and FSH regulate and stimulate the growth and function of the gonads (ovaries and testes)
synergistically.2,3
Like FSH, TSH and hCG, LH is a glycoprotein consisting of two subunits (α and ß-chains). Its
molecular weight is approx. 30,000 D. While the α-chains of all the glycoprotein hormones are very
similar, the ß-chains carry the hormone-specific information. In women the gonadotropins act within
the hypothalamus-pituitary-ovary regulating circuit to control the menstrual cycle.
Under the influence of various neurotransmitters, such as noradrenaline, dopamine and serotonin,
GnRH (gonadotropin releasing hormone) is released from the hypothalamus in pulses.
This leads to the pulsatile secretion of LH and FSH by the gonadotropic cells of the anterior pituitary
gland, with the help of cAMP (cyclic adenosine monophosphate) and calcium. (Fig. 1)
Hypothalamus
Nucleus
Noradrenaline arcuatus
LH-RH β-Endorphin
Dopamine
Oxytocin
Vasopressin
LH-RH
Portal
plexus
Gonadotropic
cells
LH, PP
FSH
FSH
AP PI
LH FSH
Inhibin
Estradiol
Progesterone Granulosa cells
Theca interna
Ovary
AP = Anterior pituitary
PI = Pars intermedia
PP = Posterior pituitary
Fig. 1: Regulatory circuits of the hypothalamus-pituitary-ovarian axis
3
LH and FSH pass via the blood-

Anterior
stream to the ovaries where they
pituitary stimulate the growth and
maturation of the follicle and
hence also the biosynthesis of
Units/l
– FSH
estrogens and progesterone.
– LH There are various feedback
mechanisms between the ovary
1234 and the hypothalamus-pituitary
1234
1234
1234
– Estradiol axis.3,4,5a,7a
1234
Progesterone
1234
– Progesterone
1234 (Fig. 2)
Estradiol
[µg/l] [µg/l]
Basal body
temperature
Increase of
Ovulation approx. 0.5°C
Maturation of
follicle in the
ovary ovum
Immature Maturing Ovu- Corpus Degener-
follicle follicle lation luteum ating cor-
pus luteum
Uterine
endometrium Fig. 2: The female menstrual cycle.
Shedding Regeneration Ischemia Cyclic changes in the blood
Days
concentrations of individual
Menstrua- Follicular phase = Luteal phase = hormones, the basal body
tion Proliferative phase Secretory phase temperature and the uterine
endometrium
The highest LH concentrations occur during the so-called mid-cycle peak (LH surge) and induce
ovulation and formation of the corpus luteum, the principal secretion product of which is
progesterone. LH stimulates the synthesis of progesterone in the corpus luteum by increasing the
transformation of cholesterol to pregnenolone. With degeneration of the corpus luteum the
progesterone concentration falls and follicle growth can be induced again, thus beginning a new
cycle. (Fig. 3)
Estradiol [pg/ml]
LH
Progesterone
Estradiol
FSH
Menstruation Ovulation Menstruation
Follicular phase Luteal phase

Proliferative phase Secretory phase
Fig. 3: Hormone profile of the menstrual cycle
4
In the Leydig cells of the testes,

Hypothalamus LH stimulates the production of
androgens, particularly of
testosterone.4,5,7 (Fig. 4)
GnRH
Pituitary gland
Inhibits LH FSH Inhibits
Induces LH
receptors
Leydig Sertoli
cells cells
Testosterone ABP Inhibin

Estradiol
Testosterone-ABP Testes
Spermatogenesis
ABP = Androgen-binding protein

Fig. 4
Clinical indication
LH-related fertility disorders can be caused both by abnormal concentrations
(hypo- or hypergonadotropic states) or by changes in the frequency of secretion of the hormone. In
addition, the concentration ratio of LH to FSH may be altered, as in the polycystic ovary syndrome
(PCO), for example, in which increased androgen production can lead to disturbance of follicle
maturation. 3,4,5
The determination of the LH concentration is used in the elucidation of functional disorders within
the hypothalamus-pituitary-gonad system. The determination of LH in conjunction with FSH is
utilized for the following indications: congenital diseases with chromosome aberrations (e.g. Turner’s
syndrome), polycystic ovaries (PCO), clarifying the causes of amenorrhea, menopausal syndrome and
suspected Leydig cell insufficiency.1,4,5,5a
Methodology
Age- and sex-related heterogeneity of the molecular structure of the proteohormone LH is described
in the literature. Thanks to the choice of antibodies, the Elecsys LH test detects all currently
described LH species. The two specific antibodies used recognize particular conformations, with the
biotinylated antibodies detecting an epitope constructed from both subunits whereas the antibody
with the ruthenium complex label detects an epitope on the ß-subunit. As a result the Elecsys LH
assay shows no cross-reactivity with FSH, TSH or hCG.
5
2.2 Elecsys Test Principle Sandwich principle. Total duration of assay: 18 minutes at 37°C.
In the first incubation step of the Elecsys LH Assay the antigen in the sample(20 µl) reacts with a
biotinylated monoclonal LH-specific antibody and a monoclonal LH-specific antibody labeled with a
ruthenium complex** to form a sandwich complex.
In the second step streptavidin-coated microparticles are added and the complex binds to the solid
phase via interaction of biotin and streptavidin. (Fig. 5)
**Tris(2,2’-bipyridyl)ruthenium(II) complex (Ru(bpy)2+

3
)
Anti-LH-Ab~Bi
SA-micro-
particle
Antigen in 9 min 9 min
sample Detection
Ru Ru Ru
Anti-LH-Ab~Ru
Fig. 5
Elecsys Measuring Cell The electrochemiluminescence measuring cell is the most important element of the entire system. It
is constructed as flow chamber and has three main tasks:
• Separation of unbound and bound substances. The streptavidin-coated microparticles laden with
immune complexes are drawn to the surface of the electrode with the help of a magnet and held
there temporarily. Unbound reagent components and excess sample material are then removed
from the measuring cell with ProCell system buffer.
• Generation of electrochemiluminescence. Application of a defined voltage induces the electro-
chemiluminescent reaction and the resulting light emission is measured directly by the
photomultiplier.
• At the end of the electrochemiluminescent reaction the microparticles are removed with
measuring cell cleaning solution (CleanCell). The measuring cell is then ready for the next
measurement. (Fig. 6)
Photomultiplier
Microparticles with
bound immuno-
complexes
Counter electrodes
Direction of
flow
Unbound
Working electrode
Ru-label
Magnet
Fig. 6
6
Electrochemilumi- The electrochemiluminescent label is a ruthenium complex (Ru(bpy) 2+ 3

). In the electrochemical
nescence Detection: reaction (Ru(bpy) 32+) is first oxidized to Ru(bpy)33+ at the surface of the electrode. At the same time
Reaction Scheme8-13 tripropylamine (TPA, component of ProCell) which is present in excess, is oxidized to form the cation
radical TPA +• which spontaneously loses a proton. The powerful oxidant Ru(bpy)33+ reacts with the
free radical TPA•, a strong reductant, to form the excited state of the ruthenium complex, (Ru(bpy) 32+).
This returns to the ground state by emitting a photon at 620 nm and is then again available for a
new light-generating reaction cycle. (Fig. 7)
•
TPA +
_ –H +
e
•
TPA TPA
_
e
Ru(bpy) 33 +
_ Ru(bpy) 32 +
e
Excited state
Ru(bpy) 32 +
Ground
state
Photon
(620 nm)
Fig. 7
Measurement A photomultiplier measures the intensity of the signal in RLU (Relative Light Units).
The voltage application phase lasts 1.4 seconds. For measurement purposes the signal produced
over a period of 0.4 seconds (0.02 – 0.42 sec.) is integrated. Part of the voltage application phase
showing this measurement window is presented diagrammatically in Fig. 8.
RLU intensity Voltage [V]
Time [sec]
Fig. 8
Evaluation The results are determined on the basis of a calibration curve. (Fig. 9)
RLU
Concentration
Fig. 9
7
3 Reagents
3.1 Contents and Elecsys LH Reagent Kit, Cat. No. 11732234 – 100 tests
Concentrations M Streptavidin-coated microparticles (transparent cap), 1 bottle, 6.5 ml:
Streptavidin-coated microparticles, 0.72 mg/ml, binding capacity: 470 ng biotin/mg
microparticles; preservative.
R1 Anti-LH Ab~biotin (gray cap), 1 bottle, 10 ml:
Biotinylated monoclonal anti-LH antibodies (mouse) 2.0 mg/l; TRIS buffer 50 mmol/l, pH 8.0;
preservative.
R2 Anti-LH Ab~Ru(bpy)32+ (black cap), 1 bottle, 10 ml:
Monoclonal anti-LH antibodies (mouse) labeled with ruthenium complex 0.3 mg/l; TRIS buffer
50 mmol/l, pH 8.0; preservative.
Calibrators* Elecsys LH CalSet II, C at. No. 03561097, for 4 x 1 ml

LH Cal1 and 2, 2 x 2 bottles for 1 ml each
LH (human) in two concentration ranges in human serum matrix.
Controls* Elecsys PreciControl Universal, Cat. No. 11731416

PreciControl Universal 1 and 2, 2 x 2 bottles for 3 ml each.
Immunoassay control on human serum basis with values in two concentration ranges.
System Reagents* Elecsys ProCell, Cat. No. 11662988, 6 x 380 ml system buffer
Elecsys ProCell M, Cat. No. 12135019, 1 x 2 l system buffer
Phosphate buffer 300 mmol/l; tripropylamine (TPA), 180 mmol/l; detergent - 0.1 %; preservatives;
pH 6.8.
Elecsys CleanCell, Cat. No. 11662970, 6 x 380 ml measuring cell cleaning solution
Elecsys CleanCell M, Cat. No. 12135027, 1 x 2 l measuring cell cleaning solution (E170)
Potassium hydroxide solution, 176 mmol/l, pH 13.2; detergent - 1%
Elecsys SysWash, Cat. No. 11930346, 1 x 500 ml washwater additive

detergent 7,5%, preservative when ready-for-use: 0,075% detergent.
Elecsys SysClean, Cat. No. 11298500, 5 x 100 ml system cleaning solution

NaOH 12%; sodium hypochlorite solution < 2% CI active
ProbeWash M, Cat. No. 03005712, 12 x 70 ml reagent needle cleaning for rinsing during operation
and when changing reagents
KOH 176 mmol/l; detergent - 1%, pH 13,2.
PreClean M, Cat. No. 03004899, 5 x 600 ml detection cleaning solution

Phosphate buffer 10 mmol/l; sodium chloride 20 mmol/l; detergent - 0.1%; preservative; pH 7.0.
Only available in the USA Elecsys CalCheck™ LH, Cat. No. 11776819, for 3 levels
LH (human) in three concentration ranges in human serum (Check 1–3).
3.2 Precautions and For in vitro diagnostic use.

Warnings Exercise the normal precautions required for handling all laboratory reagents.
Disposal of all waste material should be in accordance with local guidelines.
The calib rators and controls contain potentially infectious human material. These products have
been prepared exclusively from the blood of donors tested individually and shown by FDA approved
methods to be free from HBsAg and antibodies to HIV 1+2 and HCV.
The materials of human origin used for preparation of the calibrators have been tested for infection
with HIV, hepatitis B and hepatitis C. The results were negative.
However, as no testing method can rule out the risk of potential infection with absolute certainty the
material must be handled just as carefully as patient specimens. In the event of exposure the
directives of the responsible health authorities should be followed. 14,15
8
3.3 Reagent Handling
3.3.1 Test Reagent The reagents in the kit have been assembled into a ready-for-use unit that cannot be separated.
All information required for correct operation is read in via the respective reagent bar codes.
3.3.2 Calibrators Reconstitute the lyophilized calibrators in exactly 1.0 ml distilled water each. To use on the Elecsys
analyzers, transfer the reconstituted calibrators into appropriate labeled, empty Elecsys snap-cap
bottles. Use the calibrators for one calibration only. All data required for calibration of the LH
assay are encoded in the bar codes supplied with the calibrators, in the reagent bar code and in the
bar code on the calibrator bottle label; they are read in by scanning.
3.3.3 Controls PreciControl Universal 1 and 2 each consist of lyophilized human serum to be reconstituted with
exactly 3.0 ml distilled water. The reconstituted control is transferred to labeled empty Elecsys snap-
cap bottles. The information contained on the bar code label and on the corresponding bar code card
is read in by scanning.
3.4 Storage and Stability
3.4.1 Test Reagent Store at 2–8°C

Store the Elecsys LH reagent kits upright in order to ensure complete resuspension
of the microparticles during the automatic mixing prior to use.
Stability:
Unopened at 2–8°C: up to the stated expiration date.
After opening: twelve weeks at 2–8°C.
On the E170/Elecsys 2010: eight weeks.
On the Elecsys 1010: four weeks (stored alternately in the refrigerator
and on the analyzer – ambient temperature 20–25°C; up to
20 hours opened in total).
3.4.2 Calibrators
The lyophilized calibrators are stable up to the stated expiration date.
Stability of the reconstituted or thawed calibrators:
On the analyzer (20-25°C): use only once
At–20°C: three months (only freeze once)
3.4.3 Controls Store at 2–8°C

Stability of the lyophilized material: up to the stated expiration date.
Stability of the reconstituted control serum:
On the analyzer (20–25°C): five hours
At 20–25°C: eight hours
At 2–8°C: three days
At –20°C: one month (only freeze once)
4 Specimen Collection and Preparation
Serum collected using standard sampling tubes or tubes containing separating gel.
Plasma treated with heparin (NH4-, Li-, Na-), EDTA-K3, sodium citrate or sodium fluoride/potassium
oxalate. If sodium citrate is used the results must be corrected by +10%.
(For further information see section “Limitations - Interference”, item 5.6, page 13.)
Stable for fourteen days at 2–8°C, six months at –20°C. Only freeze once.16
When processing samples in primary tubes, follow the instructions of the tube manufacturer.
Do not use heat-inactivated samples and samples and controls stabilized with azide.
Samples containing precipitates must be centrifuged before performing the assay.
Ensure the patients samples, calibrators and controls which were stored frozen or refrigerated are at
ambient temperature (20–25°C) before measurement.
Because of possible evaporation effects, samples, calibrators and controls on the analyzer should be
measured within two hours.
Each laboratory should establish guidelines for the sampling and acceptability of the specimens.
(Specific measures may be entered in the section “Short Guide to Elecsys LH”).
9
5 Elecsys LH Testing Procedure
5.1 Materials Provided Cat. No. 1732234, Elecsys LH reagent kit for 100 tests contains:
• M Streptavidin-coated microparticles
• R1 Anti-LH Ab~biotin
• R2 Anti-LH-Ab~Ru(bpy)32+
Materials Required • Cat. No. 03561097, Elecsys LH CalSet II, for 4 x 1 ml

(but not provided) • Cat. No. 11731416, Elecsys PreciControl Universal, for 2 x 3 ml each of PreciControl
Universal 1 and 2
• General laboratory equipment
• Distilled or deionized water
• Elecsys 1010 or 2010 analyzer or MODULAR ANALYTICS E170
Accessories for Elecsys 1010 and 2010:
• Cat. No. 11662988, Elecsys ProCell, 6 x 380 ml system buffer
• Cat. No. 11662970, Elecsys CleanCell, 6 x 380 ml measuring cell cleaning solution
• Cat. No. 11776576, Elecsys CalSet Vials, empty bottles with snap-caps for calibrators
• Cat. No. 03142949, Elecsys ControlSet Vials, empty bottles with snap-caps
• Cat. No. 11930346, Elecsys SysWash, 1 x 500 ml washwater additive
• Cat. No. 11933159, Adapter for SysClean
• Cat. No. 11706829, Elecsys 1010 Assay Cup, 12 x 32 reaction vessels, or
Cat. No. 11706802, Elecsys 2010 Assay Cup, 60 x 60 reaction vessels
• Cat. No. 11706799, Elecsys 2010 Assay Tip, 30 x 120 pipette tips
Accessories for MODULAR ANALYTICS E170:
• Cat. No. 12135019, ProCell M, 1 x 2 l system buffer
• Cat. No. 12135027, CleanCell M, 1 x 2 l measuring cell cleaning solution
• Cat. No. 03023141, PC/CC-Cups, 50 cups to prewarm ProCell M and CleanCell M
• Cat. No. 03004899, PreClean M, 5 x 600 ml detection cleaning solution
• Cat. No. 03005712, ProbeWash M, 12 x 70 ml reagent needle cleaning for rinsing during
operation and when changing reagents
• Cat. No. 12102137, AssayCups/AssayTips Combimagazine M, 48 magazines x 84 reaction
vessels or pipette tips , waste bags
• Cat. No. 03023150, WasteLiner, waste bags
• Cat. No. 03027651, SysClean Adapter M
Accessories for all analyzers:
• Cat. No. 11298500, Elecsys SysClean, 5 x 100 ml system cleaning solution
Only available in the USA: Cat. No. 11776819, Elecsys LH CalCheck™, for 3 levels
5.2 Assay Procedure* For optimal performance of the assay it is important to follow the directions given for the analyzer
used, and to check that the system’s inventory of assay materials and other consumables is
adequate.
Resuspension of the microparticles takes place automatically before use. Read in of the test-specific
parameters via the respective reagent bar code. If in exceptional cases the bar code cannot be read,
enter the 15-digit sequence of numbers. The information contained in the bar codes is shown in
Fig. 10.
E170/Elecsys 2010: Bring the cooled reagents to approx. 20°C and place on the reagent disk 20°C)
of the analyzer. Avoid the formation of foam. The system automatically regulates the temperature of
the reagents and the opening/closing of the bottles.
Elecsys 1010: Bring the cooled reagents to approx. 20-25°C and place on the sample/reagent disk
(ambient temperature 20-25°C) of the analyzer. Avoid the formation of foam. The bottle caps must be
opened manually before use and closed manually after use. Store at 2-8°C after use.
10
2-D bar code Bottle bar code
Master curve Test parameter Reagent

● Test data identification
● Application data
● Calibration data
● Quality control data

● Stability data
– reagent
– calibration
Fig. 10
5.3 Calibration*
5.3.1 Standardization The Elecsys LH assay has been calibrated against the 2nd International Standard from the National
Traceability Institute for Biological Standards and Control (NIBSC) 80/552. 17
The reference standardization for the Elecsys LH master calibrators was performed by a method
comparison in 4 laboratories, with 4 series each, using the reference standard (reference material
weighed into analyte-free human serum) as reference. The calibration values determined for the five
master calibrators (minimum) give the master curve (fitting via Rodbard function) for Elecsys LH
which is stored in the reagent bar code.
The master calibrators consist of analyte-free human serum with added human LH and native human
serum samples.
Fig. 11 shows a method comparison of results based on reference standards (x) and master
calibrators (y) in 239 sera from patients and healthy subjects.
100
90
Elecsys LH – based on master calibrators [mIU/ml]
80
70
60
50
40
30
20
10
0
0 10 20 30 40 50 60 70 80 90 100
Elecsys LH – based on reference standards [mIU/ml]

Passing/Bablok 15-17 y= 0.10 + 0.99x r = 0.999
Linear regression y= -0.03 + 1.01x
Fig. 11: Reference standardization LH
11
For the Elecsys LH CalSet II reagent lot-specific reference values for 2 calibrators are determined by
calibration against the master curve. These reference values are contained in the respective bar
codes.
The CalSet calibrators consist of human serum matrix with added human LH.
The calibration on the analyzer is performed with the data from the master curve and measurement
of the two calibrators (approx. 1 mIU/ml for Cal1 und approx. 45 mIU/ml for Cal2). An example of a
typical calibration curve is shown in Fig. 12:
RLU 5.0 x 105 Typical LH calibration curve
4.0 x 105
3.0 x 105
2.0 x 105
1.0 x 105
0
0 50 100 150 200 mlU/ml
Concentration [mIU/ml]
Fig. 12
5.3.2 Calibration Calibration must be performed once per reagent lot using fresh reagent (i.e. not more than 24
Frequency* hours since the reagent pack was registered on the analyzer).
Renewed calibration is recommended as follows:
E170/Elecsys 2010
• after one month (28 days) when using the same reagent lot
• after seven days (when using the same reagent kit on the analyzer)
Elecsys 1010
• with every reagent kit
• after seven days (ambient temperature 20-25°C)
• after three days (ambient temperature 25-32°C)
For all analyzers
• as required: e.g. if quality control findings are outside the specified range.
5.3.3 Verification of Not necessary. The analyzer’s software automatically checks the validity of the curve and draws
Calibration attention to any deviations.
5.4 Quality Control* Elecsys PreciControl Universal 1 and 2. Other suitable controls can be used in addition.
Controls for the various concentration ranges should be run as single determinations at least once
every 24 hours when the test is in use, once per reagent kit and after every calibration. The control
intervals should be adapted to each laboratory’s individual requirements. Values obtained should fall
within the defined ranges.
Each laboratory should establish guidelines for corrective measures to be taken if values fall outside
the range.
(Specific measures may be entered in the section “Short Guide to Elecsys LH”)
5.5 Calculation* E170, Elecsys 1010 and 2010 automatically calculate the LH concentration of each sample either
in mIU/ml or IU/l (selectable).
12
5.6 Limitations – The LH assay is not affected by the following (criterion: recovery within ± 10 % of initial value):
Interference 17
Substance Maximum concentration No influence up to
tested concentration of
Bilirubin 66 mg/dl 66 mg/dl
Hemoglobin 1.0 g/dl 1.0 g/dl
Intralipid® 1900 mg/dl 1900 mg/dl
Biotin 100 ng/ml 50 ng/ml
In patients receiving treatment with high biotin doses (> 5 mg/day) there should be an interval of at
least 8 hours between the last biotin administration and collection of the sample.
The following commercially available plasma sample collection tubes were tested: heparin (NH+4-, Li-, Na-),
EDTA-K3, sodium citrate and sodium fluoride/potassium oxalate.
The LH concentrations were between approx. 0.1 and 60 mIU/ml.
The following table shows a comparison of serum (x) and plasma (y):
Anticoagulant Number Passing/Bablok SD (md 68)
Heparin (NH+4-, Li-, Na-) 57 y = 0.04 + 0.98x 0.07
EDTA 57 y = -0.02 + 0.97x 0.06
Citrate 56 y = 0.05 + 0.85x 0.05
Fluoride/oxalate 57 y = -0.001 + 0.92x 0.06
If sodium citrate is used the results obtained must be corrected by +10% (volume effect).
The criterion for the specimen listed in section "Specimen Collection and Preparation " are: Recovery
within 90-110% of serum value or slope 0.9–1.1 + coefficent of correlation >0.95 + intercept
within <± 2x analytical sensitivity (LDL).
No interference by rheumatoid factor up to a concentration of 1500 U/ml or in samples from dialysis
patients has been observed.
There is no high-dose hook effect at LH concentrations up to 1150 mIU/ml.
In vitro tests were performed with 17 commonly used pharmaceuticals (see table). No interference
with the LH assay was found. The LH concentrations were approx. 3 mIU/ml.
No. Active Concentration No. Active Concentration

Agent Tested [mg/l] Agent Tested [mg/l]
1 Acetylcysteine 150 10 Methyldopa 20
2 Acetylsalicylic acid 1000 11 Metronidazol 200
3 Ampicillin 1000 12 Phenylbutazone 400
4 Ascorbic acid 300 13 Rifampicin 60
5 Calcium dobesilate 20 14 Tetracycline 50
6 Cefoxitin 2500 15 Paracetamol 200
7 Cyclosporin 5 16 Ibuprofen 500
8 Soya oil (Intralipid) 10,000 17 Theophylline 100
9 Levodopa 20
Specimens from neonates have not been tested with Elecsys LH.
As with all tests containing monoclonal mouse antibodies, erroneous results may be obtained from
samples taken from patients who have been treated with monoclonal mouse antibodies or have
received them for diagnostic purposes.
In rare cases, interference due to extremely high titers of antibodies to streptavidin and ruthenium
can occur.
Elecsys LH contains additives which minimize these effects.
For diagnostic purposes, the Elecsys LH results should always be assessed in conjunction with the
patient’s medical history, clinical examination and other investigations.
13
5.7 Measuring Range 0.100–200 mIU/ml (defined by the lower detection limit and the maximum of the master curve).
(Reportable range) Values below the detection limit are reported as <0.100 mIU/ml. Values above the measuring range
are reported as >200 mIU/ml.
5.8 Dilution The broad measuring range makes dilution unnecessary.
5.9 Expected Values17,21

(Reference Range) The following LH values were obtained in studies with Elecsys LH (pooled data from MCE study
2010, March 1997, and MCE 1010, March 1998 – see also Fig. 32, p. 31).
Adults
LH (mIU/ml)
Subjects Percentile
nnnnnnn 50 5 95
Men 322 4.0 1.7 8.6
Women
• Follicular phase 316 5.9 2.4 12.6
• Ovulat. phase 56 30.8 14.0 95.6
• Luteal phase 280 4.3 1.0 11.4
• Postmenopause 132 29.1 7.7 58.5
LH/FSH quotient: Quotients were calculated from the values found with Elecsys LH and Elecsys FSH
in samples from healthy women of child-bearing age.
The following medians were obtained:
Follicular phase: 0.82 (n = 315)
Luteal phase: 1.12 (n = 279)
Children and adolescents The following tables show the results (distribution is age- and sex-dependent) of the reference
range determination for children and adolescents.
The LH concentrations were determined in samples from healthy boys and girls (age: birth to 17)
No significant sex-specific differences in LH concentration were measured in nursing infants. The
data were therefore evaluated together.
Combined data from Aachen, Hildesheim and Leipzig, D, evaluation 7/98 - see also Sect. 8.3, Fig.
32a–32b, p. 31/32.
LH (mIU/ml)
Subjects Percentile
n 50 5 95
Nursing infants- girls and boys
• < 24 hours 13 <0.1 <0.1 1.0
• 1 day 36 <0.1 <0.1 0.8
• 2 days 48 <0.1 <0.1 0.6
• 3 days 64 0.1 <0.1 2.7
• 4 days 35 <0.1 <0.1 1.7
• 5 days 25 0.5 <0.1 3.1
• 6 days 10 0.5 0.4 6.4
• 7 days 9 1.4 <0.1 5.6
• 8 - 30 days 21 2.5 <0.1 7.8
< 0.1 mIU/ml is the analytical sensitivity of the test.
14
LH (mIU/ml)
Subjects Percentile
n 50 5 95
Children and adolescents

Girls
•>1 - 12 months 14 <0.1 <0.1 0.4
• 1 - 5 years 56 <0.1 <0.1 0.5
• 6 - 10 years 58 <0.1 <0.1 3.1
• 11 - 13 years* 53 2.6 <0.1 11.9
• 14 - 17 years* 56 6.0 0.5 41.7
Boys
•>1 - 12 months 22 <0.1 <0.1 0.4
• 1 - 5 years 63 <0.1 <0.1 1.3
• 6 - 10 years 58 <0.1 <0.1 1.4
• 11 - 13 years* 23 0.9 0.1 7.8
• 14 - 17 years* 76 3.7 1.3 9.8
* There was no data available on the stage of the physical development.
Each laboratory should investigate the transferability of the expected values to its own patient
population and if necessary determine its own reference ranges. (The corresponding ranges may be
entered in the section “Short guide to Elecsys LH”.)
15
6 Specific Performance Data of the Test 17
Representative performance data for the Elecsys analyzers are given below. The results obtained in
individual laboratories may differ.
6.1 Precision Reproducibility was determined using Elecsys LH reagents (2002), pooled human sera and
controls in accordance with a modified protocol (EP5-A) of the NCCLS (National Committee for
Clinical Laboratory Standards): 6 times daily for 10 days (n = 60); within-run precision on E170, n=21.
Within-run and
Elecsys 1010 Within-run precision Total precision
Total precision
Sample Mean SD % CV SD % CV
mIU/ml mIU/ml mIU/ml
Human serum 1 0.54 0.01 1.8 0.03 5.2
Human serum 2 27.19 0.21 0.8 0.54 2.0
Human serum 3 50.72 0.41 0.8 1.01 2.0
PreciControl U 1 9.38 0.11 1.1 0.19 2.0
PreciControl U 2 44.82 0.42 0.9 0.83 1.9
Elecsys 2010 Within-run precision Total precision

Sample Mean SD % CV SD % CV
mIU/ml mIU/ml mIU/ml
Human serum 1 0.54 0.01 1.8 0.03 5.2
Human serum 2 27.19 0.21 0.8 0.54 2.0
Human serum 3 50.72 0.41 0.8 1.01 2.0
PreciControl U 1 9.38 0.11 1.1 0.19 2.0
PreciControl U 2 44.82 0.42 0.9 0.83 1.9
E170 Within-run precision Total precision

Sample Mean SD %CV Mean SD %CV
mIU/ml mIU/ml mIU/ml mIU/ml
Human serum 1 6.15 0.08 1.2 5.81 0.12 2.0
Human serum 2 92.2 0.68 0.7 89.1 1.47 1.6
Human serum 3 164 1.41 0.9 159 3.47 2.2
PreciControl U 1 6.67 0.05 0.8 6.63 0.14 2.1
PreciControl U 2 54.6 0.35 0.6 54.2 1.13 2.1
Inter-module precision The table below shows the inter-module precision results measuring Roche-internal human control
MODULAR ANALYTICS samples and control sera in 2 runs each on 4 Roche-internal MODULAR ANALYTICS E170 systems
E170
Sample Mean %CV Sample Mean %CV
mIU/ml mIU/ml
ICS 1 2.44 3.1 ICS 7 5.15 2.1
ICS 2 7.22 2.2 ICS 8 7.66 2.4
ICS 3 13.4 1.9 ICS 9 8.12 2.1
ICS 4 29.4 2.2 ICS 10 13.4 2.0
ICS 5 152 2.4 PCU 1 6.79 2.0
ICS 6 3.55 2.6 PCU 2 56.5 2.0
16
6.2 Analytical Sensitivity 0.10 mIU/ml

(Lower Detection Limit) The detection limit represents the lowest measurable LH concentration that can be distinguished
from zero. It is calculated as the concentration lying two standard deviations above the lowest
standard (master calibrator, standard 1 + 2 SD, median of three series, intra-assay precision,
n = 21).
6.3 Analytical Specificity For the monoclonal antibodies used, the following cross-reactivities were found:
Concentration Percent
used cross-reactivity
FSH 300 mIU/ml not detectable
TSH 300 µIU/ml 0.04
hCG 600 IU/ml 0.04
hGH 600 µIU/ml 0.06
hPL 13.8 pmol/ml not detectable
6.4 Method Comparisons
6.4.1 Elecsys LH/ Fig. 13 shows a comparison of the Elecsys LH method (y) with Enzymun-Test LH (x) in 166 patient
Enzymun-Test® LH sera. The results of the two tests have been converted to the restandardization of December 1997.
160
140
120
Elecsys LH, [mIU/ml]
100
80
60
40
20
0
0 20 40 60 80 100 120 140 160
Enzymun-Test LH [mIU/ml]
Passing/Bablok18--20 y = -0.46 + 1.09x SD (md 68) = 1.07 τ = 0.929

Linear regression y = -0.80 + 1.14x Sy.x = 1.72 r = 0.993
Fig. 13
17
6.4.2 Elecsys 1010/ Fig. 14 shows a method comparison between Elecsys LH on the Elecsys 1010 (y) and the Elecsys
Elecsys 2010 2010 (x) in 45 patient sera.
70
60
Elecsys LH, Elecsys 1010 [mIU/ml]
50
40
30
20
10
0
0 10 20 30 40 50 60 70
Elecsys LH, Elecsys 2010[mIU/ml]
Passing/Bablok 18--20 y = -0.02 + 0.96x SD (md 68) = 0.35 τ = 0.976

Linear regression y = 0.04 + 0.96x Sy.x = 0.49 r = 1.00
Fig. 14
6.4.3 E170/Elecsys 2010 Fig. 14a shows a method comparison between Elecsys LH on the E170 (y) and the Elecsys 2010 (x)
using 180 samples (patient sera and controls).
250
200
Elecsys LH, E170 [mIU/ml]
150
100
50
0
0 50 100 150 200 250

Passing/Bablok 18--20 y = -0.01 + 1.01x SD (md 95) = 5.02 τ = 0.969
Linear regression y = 0.10 + 1.02x Sy.x = 1.82 r = 0.999
Fig. 14a
18
6.4.4 LH CalSet II 03561097/ Fig. 14b shows the comparability of results calibrated by the new lyohilized CalSet, Elecsys LH
LH CalSet 11732269 CalSet II, 03561097 (y) - human serum matrix - compared results based on calibration with the
liquid CalSet, Elecsys LH CalSet, 11732269 (x) - bovine serum albumin matrix. The measurements
were performed on Elecsys 2010 instruments in Roche internal human control samples, human sera
and controls (n = 235).
Fig. 14c shows the evaluation of a subgroup (n = 119) in the concentration range up to 15 mIU/ml.
250 15
LH CalSet II, lyophilized, [mIU/ml]

LH CalSet II, lyophilized, [mIU/ml]
200 12
150 9
100 6
50 3
0 0
0 50 100 150 200 250 0 3 6 9 12 15
LH CalSet, liquid [mIU/ml] LH CalSet, liquid [mIU/ml]
Passing/Bablok 18–20
Fig. 14b: y= 0.03 + 0.97x SD(md 68) = 0.70 τ = 0.962
Fig. 14c: y= 0.02 + 0.98x SD(md 68) = 0.12 τ = 0.920
Linear regression Fig. 14b: y= 0.29 + 0.98x Sy.x = 2.32 r = 0.998
Fig. 14c: y= -0.01 + 0.99x Sy.x = 0.18 r = 0.997
Fig. 14b Fig. 14c
19
7 References
1. Beastall GH, Ferguson KM, O’Reilly DSJ, Seth J, Sheridan B. Assays for follicle stimulating hormone and luteinizing hormone:
Guidelines for the provision of a clinical biochemistry service. Ann Clin Biochem 1987;24:246–262.
2. Amar AP Weiss MH. Pituitary anatomy and physiology. Neurosurg Clin N Am 2003;14(1):11–23.
3. Johnson MR, Carter G, Grint C, Lightmann SL. Relationship between ovarian steroids, gonadotropin and relaxin during the
menstrual cycle. Acta Endocrinol 1993;129/2: 121–125.
4. Runnebaum B, Rabe T. Gynäkologische Endokrinologie und Fortpflanzungsmedizin Springer Verlag 1994. Band 1:17,202–
205,252–253, Band 2:350,360–362. ISBN 3–540–57345–3, ISBN 3–540–57347x.
5. Scott MG, Ladenson JH, Green ED, Gast MJ. Hormonal evaluation of female infertility and reproductive disorders. Clin Chem
1989;35:620–630.
5a. Pelinck MJ, Hoek A, Simons AH, Heineman MJ. Efficacy of natural cycle IVF: a review of the literature. Hum Reprod Update
2002;8(2):129–139.
6. Van Ginkel LA, Loeber LG. Heterogeneity of human luteinizing hormone. Discrimination between acidic and basic preparations.
Acta Endocrinol 1989;121:73–82.
7. Juhl UM, Rippegather G, Weller J, Zawta B. Important Facts on Reproduction Medicine/Fertility Diagnosis, Questions and
Answers. 1994. Boehringer Mannheim, Cat.-No. 1322958.
7a. Park SJ, Goldsmith LT, Weiss G. Age-related changes in the regulation of luteinizing hormaone secretion by estrogen in women.
Exp Biol Med 2002;227(7):455–64.
8. Blackburn GF et al. Electrochemiluminescence Detection for Development of Immunoassays and DNA Probe Assays for Clinical
Diagnostics. Clin Chem 1991;37(9):1534–39.
9. Kenten JH et al. Rapid Electrochemiluminescence Assays of Polymerase Chain Reaction Products. Clin Chem 1991;37(9):
1626–32.
10. Kenten JH et al. Improved Electrochemiluminescent label for DNA Probe Assays: Rapid Quantitative Assays of HIV-1 Polymerase
Chain Reaction Products. Clin Chem 1992;38(6):873–79.
11. Leland JK, Powell MJ. Electrogenerated Chemiluminescence: An Oxidative-Reduction Type ECL Reaction/Sequence Using
Tripropyl Amine. J Electrochem Soc 1990;137(10):3127–31.
12. Obeng YS, Bard AJ. Electrogenerated Chemiluminescence. 53 Electrochemistry and Emission from Adsorbed Monolayers of a
Tris (bipyridyl) ruthenium (II)-Based Surfactant on Gold and Tin Oxide Electrodes. Langmuir 1991;7:195–201.
13. Xu X-H, Bard AJ. Electrogenerated Chemiluminescence. 55 Emission from Adsorbed Ru(bpy)2+
3
on Graphite, Platinum, and Gold.
Langmuir 1994;10:2409–2414.
13. Occupational Safety and Health Standards: bloodborne pathogens (29 CFR Part 1910.1030 - Occupational Safety and Fed.
Register July 1, 2001;17: 260-273.
14. Council Directive (2000/54EC). Official Journal of the European Communities No. L262 from Oct. 17,2000.
16. Tietz NW. Clinical Guide To Laboratory Tests. 3rd ed. Philadelphia, Pa: WB Saunders Co, 1995:410.
17. Data on file at Roche Diagnostics
18. Passing H, Bablok W. A New Biometrical Procedure for Testing the Equality of Measurements from Two Different Analytical
Methods. J Clin Chem Clin Biochem 1983;21:709–20.
19. Passing H, Bablok W. Comparison of Several Regression Procedures for Method Comparison Studies and Determination of
Sample Sizes. J Clin Chem Clin Biochem 1984;22:431–445.
20. Bablok W et al. A General Regression Procedure for Method Transformation. J Clin Chem Clin Biochem 1988;26:783-790.
21. Gassler N, Peuschel T, Pankau R. Pediatric reference values of estradiol, testosterone, lutropin, follitropin and prolactin. Clin
Lav 2000; 46(11-12):553–560
* For further information, please consult the operators’ manuals for Elecsys 1010/2010/MODULAR ANALYTICS E170, as well as
the package inserts for the system reagents, LH CalSet II and PreciControl Universal.
Manufactured by Roche Diagnostics GmbH, Germany.

Elecsys®, Enzymun-Test® and COBAS® are registered trademarks of Roche Diagnostics GmbH.
CalCheck™ is a trademark of Roche Diagnostics Corporation.
All other trademarks are the property of their respective holders.
20
8 Results of Multicenter Studies
Introduction This section is intended to provide information on external studies. The data presented below are
derived from the following studies:
• Multicenter study “Elecsys 2010 Fertility” at 10 clinical centers in different countries; data
presented in March 1997. The data will be referred to as “Data: MCE Elecsys 2010, 3/97”.
• Multicenter study “Elecsys 1010 Fertility” at 10 clinical centers in different countries; data
summarized in January 1998. The data will be referred to as “Data: MCE Elecsys 1010, 1/98”.
NB : All Elecsys LH and Enzymun-Test® LH results in the method comparisons have been converted to
the restandardization of December 1997. This means that there are differences with respect to the
method comparisons contained in the reports.
• Reference ranges for children were determined using data obtained in Aachen, Hiltdesheim, and
Leipzig. (Evaluation 7/98).
• Multicenterstudy „MODULAR ANALYTICS <EE>“ (2 E170-modules), in 9 clinical centers in different
countries; data presented in September 2001. The data will be referred to as: „MCE MODULAR
ANALYTICS <EE>, 9/2001”.
8.1 Precision
8.1.1 Intra-assay precision Fig. 15: Eight laboratories (Elecsys 2010) and 4 laboratories (Elecsys 1010) each measured 3 human
sera of different concentration ranges in one series of 21 determinations on one day and 5
laboratories each measured 2 human sera and 2 controls on MODULAR ANALYTICS E170.
20 MCE Elecsys 1010

MCE Elecsys 2010
18
MCE E170
16
14
12
10
CV [%]
8
6
4
2
0
1 10 100 1000
LH [mIU/ml]
Fig. 15: MCE 2010, 3/97 + MCE 1010, 1/98 + MCE MODULAR ANALYTICS <EE> 9/2001
8.1.2 Interassay precision Fig. 16: Seven laboratories (Elecsys 2010) and 4 laboratories (Elecsys 1010) each measured 3
Total precision human sera of different concentration ranges as single determinations on 10–21 days and 5
laboratories each measured 2 human sera and 2 controls on MODULAR ANALYTICS E170 (total
precision).
20 MCE Elecsys 1010

18 MCE Elecsys 2010
MCE E170
16
14
12
CV [%]
10
0
1 10 100 1000
LH [mIU/ml]
Fig. 16: MCE 2010, 3/97 + MCE 1010, 1/98 + MCE MODULAR ANALYTICS <EE> 9/2001
21
8.1.3 Inter-module (E170)/ Figure 17 displays the inter-module precision data in human serum samples, controls measured on
inter-instrument MODULAR ANALYTICS <EE> (2 E170 modules) during the Multicenter Evaluation and the data from
precision the table, page 16.
10 MCE <EE>
MODULAR ANALYTICS E170

8
6
CV [%]
0
1 10 100 1000
LH [mIU/ml]
Fig.17: MCE MODULAR ANALYTICS <EE> 9/2001 + Roche interne Daten
8.1.4 Inter-Instrument The inter-instrument precision performed in Roche-internal human contols sampels, human sera,
precision - calibration and controls in 2 series each on 4 Elecsys 1010/2010 and MODULAR ANALYTICS E170 is shown in
with LH CalSet II Fig. 18. The results were obtained using 2 different lots of LH CalSet II and test reagents.
10 LH CalSet II lot no. 1,

Elecsys 1010 Reagent lot no.162252
8 LH CalSet II lot 165288,
Reagent Ch. 165260
6
CV [%]
0
1 10 100 1000

Elecsys 2010 Reagent lot no.162252
Reagent Ch. 165260
CV [%]
0
1 10 100 1000

E170
Reagent lot no.162252
Reagent Ch. 165260
CV [%]
0
1 10 100 1000
LH [mIU/ml]
Fig.18: Roche-internal data - 6/'03
22
8.2 Method comparisons
8.2.1 E170/ Fig. 19 shows a method comparison between Elecsys LH on MODULAR ANALYTICS E170 (y) and on the
Elecsys 2010 Elecsys 2010 (x) in 138 patient sera (MCE MODULAR ANALYTICS <EE> 9/2001, Balitmore, USA).
Fig. 19a shows the evaluation of a subgroup (n = 108) in the concentration range up to 20 mIU/ml.
200 20
Elecsys LH, E 170 [mIU/ml]

160 16
120 12
80 8
40 4
0 0
0 40 80 120 160 200 0 4 8 12 16 20
Elecsys 2010, Elecsys LH [mIU/ml] Elecsys 2010, Elecsys LH [mIU/ml]
Passing/Bablok 18–20
Fig. 19: y = 0.04 + 1.03x SD(md 68) = 0.27 τ = 0.976
Fig. 19a: y = 0.04 + 1.03x SD(md 68) = 0.19 τ = 0.967
Linear regression Fig. 19: y = -0.02 + 1.05x Sy.x = 0.49 r = 0.999
Fig. 19a: y = 0.04 + 1.04x Sy.x = 0.26 r = 0.998
Fig. 19 Fig. 19a
8.2.2 Elecsys 1010/ Fig. 20 shows a method comparison between Elecsys LH on the Elecsys 1010 (y) and on the Elecsys 2010
Elecsys 2010 (x) in 158 patient sera (MCE 1010, 1/98, Heidelberg, D).
60
50
40
30
20
10
0
0 10 20 30 40 50 60
Passing/Bablok 18-- 20 y= -0.05 + 1.01x SD (md 68) = 0.22 r = 0.997

Linear regression y= -0.04 + 1.04x Sy.x = 0.71
Fig. 20: Data: MCE Elecsys 1010, 1/98
23
8.2.3 Elecsys LH The following two method comparisons between Elecsys LH on MODULAR ANALYTICS E170 and
LH ADVIA® Centaur ® the Bayer LH test on the ADVIA Centaur were performed in 2 clinical centers (MCE MODULAR
ANALYTICS <EE> 9/2001).
Fig. 21 shows a method comparison between Elecsys LH on MODULAR ANALYTICS E170 (y) and the
Bayer LH test on the ADVIA Centaur (x) in 150 patient sera (MCE MODULAR ANALYTICS <EE> 9/2001,
Augsburg, D).
Fig. 21a shows the evaluation of a subgroup (n = 124) in the concentration range up to 20 mIU/ml.
200 20

160 16
120 12
80 8
40 4
0 0
0 40 80 120 160 200 0 4 8 12 16 20
LH ADVIA Centaur [mIU/ml] LH ADVIA Centaur [mIU/ml]
Passing/Bablok18–20 Fig. 21: y= 0.44 + 0.96x SD(md 68) = 0.42 τ = 0.931

Fig. 21a: y= 0.41 + 0.97x SD(md 68) = 0.30 τ = 0.907
Linear regression Fig. 21: y= 0.87 + 0.93x Sy.x = 1.10 r = 0.991
Fig. 21a: y= 0.51 + 0.97x Sy.x = 0.50 r = 0.987
Fig. 21 Fig. 21a
Fig. 21b shows a method comparison between Elecsys LH on MODULAR ANALYTICS E170 (y) and the
Bayer LH test on the ADVIA Centaur (x) in 149 patient sera (MCE MODULAR ANALYTICS <EE> 9/2001,
Heidelberg, D).
Fig. 21c shows the evaluation of a subgroup (n = 104) in the concentration range up to 20 mIU/ml.
200 20
160 16
120 12
80 8
40 4
0 0
0 40 80 120 160 200 0 4 8 12 16 20
LH ADVIA Centaur [mIU/ml] LH ADVIA Centaur [mIU/ml]
Passing/Bablok18–20 Fig. 21b: y= 0.35 + 0.95x SD(md 68) = 0.87 τ = 0.919

Fig. 21c: y= 0.14 + 1.00x SD(md 68) = 0.45 τ = 0.870
Linear regression Fig. 21b: y= 1.35 + 0.91x Sy.x = 1.65 r = 0.994
Fig. 21c: y= 0.33 + 1.04x Sy.x = 1.13 r = 0.932
Fig. 21b Fig. 21c
24
8.2.4 Elecsys LH/ Fig. 22 shows a method comparison between Elecsys LH on the Elecsys 2010 (y) and the TOSOH LH test
LH TOSOH AIA 1200 on the AIA 1200 (x) in 170 patient sera (MCE 2010 3/97, Tokyo, J).
140
120
Elecsys 2010, Elecsys LH [mIU/ml]
100
80
60
40
20
0
0 20 40 60 80 100 120 140
AIA 1200 LH [mIU/ml]
Passing/Bablok 18-- 20 y= 0.51 + 1.08x SD (md 68) = 0.64 r = 0.989
Linear regression y= 0.95 + 1.00x Sy.x = 1.78
8.2.5 Elecsys LH/ Fig. 23 shows a method comparison between Elecsys LH on the Elecsys 2010 (y) and the bioMérieux LH
LH bioMérieux test (IRMA) (x) in 121 patient sera (MCE 2010 3/97, Brussels, B).
70
60
50
40
30
20
10
0
0 10 20 30 40 50 60 70
LH RIA bioMérieux [mIU/ml]

25
8.2.6 Elecsys LH/ Fig. 24 shows a method comparison between Elecsys LH on the Elecsys 2010 (y) and the Pharmacia
LH Pharmacia Wallac LH Test (RIA) (x) in 128 patient sera (MCE 2010, 3/97, Leiden, NL).
100
90
80
70
60
50
40
30
20
10
0
0 10 20 30 40 50 60 70 80 90 100
LH Pharmacia [mIU/ml]
Passing/Bablok18-- 20 y= 0.22 + 1.19x SD (md 68) = 1.06 r = 0.986
8.2.7 Elecsys LH/ Fig. 25 shows a method comparison between Elecsys LH on the Elecsys 1010 (y) and the Roche
LH Cobas Core® Diagnostics LH test (x) on the Cobas Core in 242 patient sera (MCE 1010 1/98, Basle, CH)
70
60
50
40
30
20
10
0
0 10 20 30 40 50 60 70
LH Cobas Core [mIU/ml]

Passing/Bablok 18-- 20 y = -0.27 + 1.11x SD (md 68) = 0.51 r = 0.987
Linear regression y = -0.24 + 1.08x Sy.x =1.06
Fig. 25: Data MCE Elecsys 1010, 1/98
26
8.2.8 Elecsys LH/ Fig. 26 shows a method comparison between Elecsys LH on the Elecsys 1010 (y) and the Abbott LH
LH AxSym® test (x) on the AxSym in 104 patient sera (MCE 1010 1/98, Chicago, USA)
.
140
120
100
80
60
40
20
0
0 20 40 60 80 100 120 140
LH AxSym [mIU/ml]
8.2.9 Elecsys LH/ Fig. 27 shows a method comparison between Elecsys LH on the Elecsys 1010 (y) and the Bayer LH
LH Bayer Immuno 1 test (x) on the Immuno 1 in 37 patient sera (MCE 1010 1/98, Heidelberg, D).
60
50
40
30
20
10
0
0 10 20 30 40 50 60
LH Immuno 1 [mIU/ml]
Passing/Bablok18-- 20 y= 0.33 + 0.96x SD (md 68) = 0.29 r = 0.99
27
8.2.10 Elecsys LH/ Fig. 28 shows a method comparison between Elecsys LH on the Elecsys 1010 (y) and the Bayer LH
LH ACS: 180 test on the ACS: 180 (x) in 107 patient sera (Heidelberg, D).
60
50
40
30
20
10
0
0 10 20 30 40 50 60
LH ACS:180 [mIU/ml]
8.2.11 Elecsys LH/ Fig. 29 shows a method comparison between Elecsys LH on the Elecsys 1010 (y) and the Serono LH test
LH Serono (RIA) (x) in 155 patient sera (MCE 1010 1/98, Giessen, D).
14
12
10
0
0 2 4 6 8 10 12 14
LH Serono [mIU/ml]
Passing/Bablok 18- 20 y = 0.15 + 1.36x SD (md 68) = 0.50 r = 0.92 Linear
regression y = 0.56 + 1.23x Sy.x = 0.56
28
8.2.12 Elecsys LH The following two method comparisons were performed after restandardisation of Elecsys LH
in the same specimen panel in December 1997 (Penzberg, D).
Elecsys LH/ Fig. 30 shows a method comparison between Elecsys LH on the Elecsys 2010 (x) and the Bayer LH
LH ACS:180 test on the ACS: 180 (y) in 91 patient sera.
140
120
100
LH ACS:180 [mIU/ml]
80
60
40
20
0
0 20 40 60 80 100 120 140
Passing/Bablok18-- 20 y= -0.06 + 0.85x r = 0.988
Fig. 30: Data, 12/97
Elecsys LH/ Fig. 31 shows a method comparison between Elecsys LH on the Elecsys 2010 (x) and the Abbott LH
LH IMx® test on the IMx (y) in 104 patient sera.
140
120
100
LH IMx [mIU/ml]
80
60
40
20
0
0 20 40 60 80 100 120 140
18-- 20
Passing/Bablok y= -0.10 +1.17x r = 0.986
Fig. 31: Data, 12/97
29
8.3 Clinical data –

Reference values
Adults Fig. 32 shows the reference ranges determined in the course of the MCE study Elecsys 2010, March
1997 and Elecsys 1010. LH concentrations were determined in samples from healthy men
andwomen of reproductive age together with further fertility parameters. The results were evaluated
together in March ’98. The figure presents graphically the individual results given in the tables in
section 5.9 and in the Short Guide.
LH/FSH Quotient : Quotients were calculated from the values found with Elecsys LH and Elecsys FSH
in samples from healthy women of child-bearing age.
The following medians were obtained (not shown in figure):
Follicular phase: 0.82 (n = 315)
Luteal phase: 1.12 (n = 279)
Men Women
Ovulat. Postm.-
Follicular phase phase Luteal phase pause
1000
100
LH [mlU/ml]
10
0.1
1 dot =
´ ` 3 values 5–95% percentile range
Fig. 32
Children and Adolescents The results (broken down by age and sex) of the reference range determination for children and
adolescents are shown in Fig. 32a-32b on page 31.
The concentration of LH was determined in samples obtained from healthy boys and girls (age: birth
to 17). No information was available regarding the stage of physical development.
No significant sex-specific differences in LH concentration were measured in nursing infants. The
data were therefore evaluated together.
The data obtained in Aachen, Hildesheim, and Leipzig, all D, were combined (Evaluation, July 1998).
Refer also to Sect. 5.9, pages 14/15.
30
Girls
Stated age: 15 Years
70
14 n = 56 n = 58 n = 53 n = 56
60
50
41.7
LH [mIU/ml]
40
30
20
11.9
10
3.1
0.4 0.5
0
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19
Age [Years] 5–95% percentile range
Fig 32a: Reference range for young girls – Data: 7/98
Boys
Stated age: 15 Years
12
22 n = 63 n = 58 n = 23 n = 76
10 9.8
8 7.8
LH [mIU/ml]
2
0.4 1.3 1.4
0
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19
Age [Years] 5–95% percentile range
Fig. 32b: Reference range for young boys – Data: 7/98
Hormone profiles Figs. 33–35, page 32, show examples of the hormone profiles of LH, FSH, estradiol and progesterone
in typical menstrual cycles:
Fig. 33: Normal cycle of a healthy woman
Fig. 34: Cycle in a woman on contraceptives
Fig. 35: Cycle after stimulation for in vitro fertilization treatment
31
40 16
35 14
30 12
LH [mlU/ml]
LH [mlU/ml]
25 10
20 8
15 6
10 4
5
2
0 0
14
9
12 8
10 7
FSH [mlU/ml]
6
FSH [mlU/ml]
8
5
6 4
4 3
2
2
1
0
0
900 2500
800
Estradiol [pmol/l]
2000
Estradiol [pmol/l]
700
600
1500
500
400 1000
300
200 500
100
0 0
45 10
Progesterone [nmol/l]
40
35 8
30
25 6
20
4
15
10
2
5
0 0
0 5 10 15 20 25 30 35 0 5 10 15 20 25 30
Day of cycle Day of cycle
Fig. 33: Normal cycle of a healthy woman Fig. 34: Cycle in a woman on contraceptives
1. 2.3. 1. 2.3.
○
○
4.00 6000
Estradiol [pmol/
○
○
3.50 5000
LH [mlU/ml]
3.00
○
○
2.50 4000
○
○
2.00 3000
○
○
1.50 2000
○
○
1.00
1000
○
○
0.50
0.00 0
○
○
○
○
250.0
○
25
○
○
○
20 200.0
FSH [mlU/ml]
○
○
150.0
○
○
15
○
○
Stimulation with FSH 100.0

10 Stimulation with FSH
○
○
Blockade of pituitary 50.0

○
○
5 Blockade of pituitary
○
0.0
○
18.10.
25.10.
01.11.
06.11.
12.11.
13.11.
17.11.
19.11.
21.11.
22.11.
23.11.
29.11.
02.12.
05.12.
08.12.
11.12.
○
0
18.10.
25.10.
01.11.
06.11.
12.11.
13.11.
17.11.
19.11.
21.11.
22.11.
23.11.
29.11.
02.12.
05.12.
08.12.
11.12.
Date Date
1. Ovulation
○ ○ ○ ○ 2. Removal of follicle and endoplasmic injection of sperm
3. Embryo returned to uterus
Fig. 35: Cycle after stimulation for in vitro fertilisation treatment
32
Notice
33
Notice
34
Notice
35
9 Short Guide to Elecsys LH
Intended Use Immunoassay for the in vitro quantitative determination of LH in human serum and plasma.
Reagents Store the Elecsys LH reagent kit upright at 2–8°C.

The test reagents are ready for use.
Bring the cooled reagents to 20°C (Elecsys 2010) or 20–25°C (Elecsys 1010) before placing them on
the reagent disk of the analyzer.
Avoid the formation of foam.
Stability:
Unopened at 2–8°C: up to the stated expiration date.
After opening: twelve weeks at 2–8°C.
On the E170/Elecsys 2010: eight weeks.
On the Elecsys 1010: four weeks (stored alternately in the refrigerator and on the analyzer –
ambient temperature 20–25°C; up to 20 hours opened in total).
Specimen Collection Serum and plasma treated with heparin (NH4-, Li-, Na-), EDTA-K3, sodium citrate or sodium fluoride/
and Preparation potassium oxalate. If sodium citrate is used the results must be corrected +10 %.
Stable for fourteen days at 2–8°C, six months at –20°C. Only freeze once.
No interference by: hemoglobin up to 1.0 g/dl, bilirubin up to 66 mg/dl, Intralipid® up to 1900 mg/dl,
biotin up to 50 ng/ml, rheumatoid factors up to 1500 U/ml.
Do not use heat-inactivated samples and samples and controls stabilized with azide.
Samples containing precipitates must be centrifuged before performing the assay.
Ensure the patients samples, calibrators and controls which were stored frozen or refrigerated are at
ambient temperature (20–25°C) before measurement.
Because of possible evaporation effects, samples, calibrators and controls on the analyzer should be
measured within two hours.
(For further information see section “Limitations – Interference”).
The following steps are to be taken if results from a sample are considered questionable:
Assay Procedure All instrument- and test-specific parameters are read in via the respective bar codes.
Calibrators 2-point calibration with LH CalSet II. Reconstitute the lyophilized calibrators and transfer them to
appropriately labeled Elecsys snap-cap bottles.
Stability of the reconstituted calibrators (freeze immediately):
At –20°C: three months (freeze only once)
On the anlyzer (20–25°C): Use only once
Calibration Frequency Calibration must be performed once per reagent lot using fresh reagent (i.e. not more than 24 hours
since the reagent pack was registered on the analyzer).
Renewed calibration is recommended as follows:
E170/Elecsys 2010
• after one month (28 days) when using the same reagent lot
• after seven days (when using the same reagent kit on the analyzer)
Elecsys 1010
• with every reagent kit
• after seven days (ambient temperature 20–25°C)
• after three days (ambient temperature 25–32°C)
For all analyzers
• whenever additionally required (e.g. if quality control findings are outside the specified range).
The following measures are to be taken if results are considered questionable:
Quality Control Elecsys PreciControl Universal 1 and 2. Controls in two concentration ranges.
The target values and ranges are contained in the bar codes. Other suitable controls can be used in
addition; enter target values manually.
36
Control Intervals Controls for the various concentration ranges should be run as single determinations at least once
every 24 hours when the test is in use, once per reagent kit and after every calibration.
The control intervals should be adapted to each laboratory’s individual requirements.
Controls must be run at the following intervals:
The following corrective measures should be taken if values fall outside the specified range
(examples):
• Run the control again with the same and a newly reconstituted control.
• Run the control again following renewed calibration.
• Run the control again using a new reagent kit.
• Repeat assays on samples giving questionable results and compare with original values.
• Check that the analyzer is working properly.
Other laboratory-specific measures:
Expected Values Reference values (mIU/ml; 5th–95th percentile) determined with Elecsys LH:
(Reference Range)
Men 1,7 – 8,6
Women
• Follicular phase 2,4 – 12,6
• Ovulation phase 14,0 – 95,6
• Luteal phase 1,0 – 11,4
• Postmenopause 7,7 – 58,5
LH/FSH quotient (median): follicular phase: 0.82 luteal phase: 1.12

Reference values for children see Sect. 5.9 u. 8.3, pages 14/15, 30/31
Dilution The broad measuring range makes dilution unnecessary.
Measuring Range/ Between lower detection limit and maximum of master curve: 0.100 to 200 mIU/ml. Samples
Reportable Range with concentrations below the detection limit of 0.100 mIU/ml are reported as <0.100 mIU/ml.
Samples with concentrations above the measuring range are reported as >200 mIU/ml.
Laboratory-specific Reference Ranges
Critical Values
Measures to be taken if critical values occur
Back-up Method
Confirmatory Tests
Checked by Name Name

Function Function
Date Date
Name Name
Function Function
Date Date
37
Elecsys LH – Overview Name of test Elecsys LH

Method ECLIA, sandwich principle
Duration of test; temperature 18 minutes, 37°C
Evaluation Quantitative
Test reagent Ready for use
Reagent volumes (M/R1/R2) 30/75/75 µl
Stability on the analyzer 8 weeks (E170/Elecsys 2010)
Calibrators LH CalSet II, 2 calibrators, lyophilized
Calibrator determination 2-fold
Calibration frequency E170/Elecsys 2010:

once per reagent lot and/or once per month or
every 7 days
Elecsys 1010:
once per reagent kit and once every 7 or 3 days
Validity of lot calibration 1 month (E170/Elecsys 2010)
Controls PreciControl Universal, lyophilized
Sample material Serum; plasma (NH4-, Li-, Na-heparin, EDTA, citrate,

fluoride/oxalate)
Sample volume 20 µl
Sample determination Single
Sample dilution –
Automatic dilution Not necessary
Sample diluent –
Sample pretreatment No
Measuring/reportable range 0.100 – 200 mIU/ml
Dynamic range 0.100 – 200 mIU/ml
Sensitivity (analytical) 0.100 mIU/ml
Expected values Men* 1.7 – 8.6

(5–95% Percentile; mIU/ml) Women* • Follicular phase 2.4 – 12.6
*More details and results for • Ovulation phase 14.0 – 95.6
nursing infants see p. 14/15. 30/31. • Luteal phase 1.0 – 11.4
• Postmenopause 7.7 – 58.5
Girls* • >1 - 12 months <0.1 – 0.4
• 1 - 5 years <0.1 – 0.5
• 6 - 10 years <0.1 – 3.1
• 11 - 13 years* <0.1 – 11.9
• 14 - 17 years* 0.5 – 41.7
Boys* • >1 - 12 months <0.1 – 0.4
06.2003 1 845 527 º USA: 011602900-0603
• 1 - 5 years <0.1 – 1.3

• 6 - 10 years <0.1 – 1.4
• 11 - 13 years* 0.1 – 7.8
June 2003 · © Roche Diagnostics
• 14 - 17 years* 1.3 – 9.8
Roche Diagnostics GmbH Roche Diagnostics Corporation

D-68298 Mannheim Indianapolis, IN, USA
Germany US Costomer Technical Support
38 1-800-428-2336

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Elecsys® 1010/2010/MODULAR ANALYTICS E170

Product Information Elecsys® LH Immunoassay

Elecsys® Systems 1010/2010

2.1 Biochemistry, Clinical Indications, Methodology 3–5

3.1 Contents and Concentrations 8

4 Specimen Collection and Preparation 10

5 Elecsys LH Testing Procedure 10

5.1 Materials Provided, Materials Required (but not provided) 10

6 Specific Performance Data of the Test 16

6.1 Precision, Within-run / Total / Inter-Instrument / Inter-Module Precision 16

8 Results of Multicenter Studies 21

9 Short Guide to Elecsys LH

2.1 Biochemistry, Biochemistry

Fig. 1: Regulatory circuits of the hypothalamus-pituitary-ovarian axis

LH and FSH pass via the blood-

Menstruation Ovulation Menstruation

Follicular phase Luteal phase

Fig. 3: Hormone profile of the menstrual cycle

In the Leydig cells of the testes,

Inhibits LH FSH Inhibits

Testosterone ABP Inhibin

ABP = Androgen-binding protein

**Tris(2,2’-bipyridyl)ruthenium(II) complex (Ru(bpy)2+

Electrochemilumi- The electrochemiluminescent label is a ruthenium complex (Ru(bpy) 2+ 3

RLU intensity Voltage [V]

Calibrators* Elecsys LH CalSet II, C at. No. 03561097, for 4 x 1 ml

Controls* Elecsys PreciControl Universal, Cat. No. 11731416

Elecsys SysWash, Cat. No. 11930346, 1 x 500 ml washwater additive

Elecsys SysClean, Cat. No. 11298500, 5 x 100 ml system cleaning solution

PreClean M, Cat. No. 03004899, 5 x 600 ml detection cleaning solution

3.2 Precautions and For in vitro diagnostic use.

3.3 Reagent Handling

3.4 Storage and Stability

3.4.1 Test Reagent Store at 2–8°C

3.4.3 Controls Store at 2–8°C

4 Specimen Collection and Preparation

5 Elecsys LH Testing Procedure

Materials Required • Cat. No. 03561097, Elecsys LH CalSet II, for 4 x 1 ml

2-D bar code Bottle bar code

Master curve Test parameter Reagent

● Quality control data

Elecsys LH – based on reference standards [mIU/ml]

Fig. 11: Reference standardization LH

RLU 5.0 x 105 Typical LH calibration curve

Bilirubin 66 mg/dl 66 mg/dl

Hemoglobin 1.0 g/dl 1.0 g/dl

Intralipid® 1900 mg/dl 1900 mg/dl

Biotin 100 ng/ml 50 ng/ml

Anticoagulant Number Passing/Bablok SD (md 68)

Heparin (NH+4-, Li-, Na-) 57 y = 0.04 + 0.98x 0.07

EDTA 57 y = -0.02 + 0.97x 0.06

Citrate 56 y = 0.05 + 0.85x 0.05

Fluoride/oxalate 57 y = -0.001 + 0.92x 0.06

No. Active Concentration No. Active Concentration

5.8 Dilution The broad measuring range makes dilution unnecessary.

5.9 Expected Values17,21

Nursing infants- girls and boys

• < 24 hours 13 <0.1 <0.1 1.0

• 1 day 36 <0.1 <0.1 0.8