ms_03737551190V15.

Ferritin
Ferritin

Elecsys 2010
MODULAR ANALYTICS E170
03737551 190 100 cobas e 411
cobas e 601
cobas e 602

English ▪ The reaction mixture is aspirated into the measuring cell where the
microparticles are magnetically captured onto the surface of the
Intended use electrode. Unbound substances are then removed with
Immunoassay for the in vitro quantitative determination of ferritin in human ProCell/ProCell M. Application of a voltage to the electrode then induces
serum and plasma. chemiluminescent emission which is measured by a photomultiplier.
The electrochemiluminescence immunoassay “ECLIA” is intended for use ▪ Results are determined via a calibration curve which is instrument-
on Elecsys and cobas e immunoassay analyzers. specifically generated by 2‑point calibration and a master curve provided
Summary via the reagent barcode.
Ferritin is a macromolecule with a molecular weight of at least 440 kDa a) Tris(2,2'-bipyridyl)ruthenium(II)-complex (Ru(bpy) )
(depending on the iron content) and consists of a protein shell (apoferritin) Reagents - working solutions
of 24 subunits and an iron core containing an average of approximately The reagent rackpack is labeled as FERR.
2500 Fe3+ ions (in liver and spleen ferritin).1
Ferritin tends to form oligomers, and when it is present in excess in the cells M Streptavidin-coated microparticles (transparent cap), 1 bottle, 6.5 mL:
of the storage organs there is a tendency for condensation to
semicrystalline hemosiderin to occur in the lysosomes. Streptavidin-coated microparticles 0.72 mg/mL; preservative.
At least 20 isoferritins can be distinguished with the aid of isoelectric R1 Anti‑Ferritin‑Ab~biotin (gray cap), 1 bottle, 10 mL:
focusing.2 This microheterogeneity is due to differences in the contents of Biotinylated monoclonal anti‑ferritin antibody (mouse) 3.0 mg/L;
the acidic H and weakly basic L subunits. The basic isoferritins are
responsible for the long‑term iron storage function, and are found mainly in phosphate buffer 100 mmol/L, pH 7.2; preservative.
the liver, spleen, and bone marrow.1,3 R2 Anti‑ferritin‑Ab~Ru(bpy) (black cap), 1 bottle, 10 mL:
Acidic isoferritins are found mainly in the myocardium, placenta, and tumor
tissue. They have a lower iron content and presumably function as Monoclonal anti‑ferritin antibody (mouse) labeled with ruthenium
intermediaries for the transfer of iron in various syntheses.4,5,6 complex 6.0 mg/L; phosphate buffer 100 mmol/L, pH 7.2;
The determination of ferritin is a suitable method for ascertaining the iron preservative.
metabolism situation. Determination of ferritin at the beginning of therapy
provides a representative measure of the body's iron reserves. A storage Precautions and warnings
deficiency in the reticulo-endothelial system (RES) can be detected at a For in vitro diagnostic use.
very early stage.7 Exercise the normal precautions required for handling all laboratory
reagents.
Clinically, a threshold value of 20 µg/L (ng/mL) has proved useful in the Disposal of all waste material should be in accordance with local guidelines.
detection of prelatent iron deficiency. This value provides a reliable Safety data sheet available for professional user on request.
indication of exhaustion of the iron reserves that can be mobilized for
hemoglobin synthesis. Latent iron deficiency is defined as a fall below the Avoid foam formation in all reagents and sample types (specimens,
12 µg/L (ng/mL) ferritin threshold. These two values necessitate no further calibrators and controls).
laboratory elucidation, even when the blood picture is still morphologically Reagent handling
normal. If the depressed ferritin level is accompanied by hypochromic,
microcytal anemia, then manifest iron deficiency is present.1 The reagents in the kit have been assembled into a ready‑for‑use unit that
cannot be separated.
When the ferritin level is elevated and the possibility of a distribution
disorder can be ruled out, this is a manifestation of iron overloading in the All information required for correct operation is read in from the respective
body. 400 µg/L (ng/mL) ferritin is used as the threshold value. Elevated reagent barcodes.
ferritin values are also encountered with the following tumors: acute Storage and stability
leukemia, Hodgkin's disease and carcinoma of the lung, colon, liver and Store at 2‑8 °C.
prostate. The determination of ferritin has proved to be of value in liver
metastasis. Studies indicate that 76 % of all patients with liver metastasis Do not freeze.
have ferritin values above 400 µg/L (ng/mL). Reasons for the elevated Store the Elecsys reagent kit upright in order to ensure complete
values could be cell necrosis, blocked erythropoiesis or increased synthesis availability of the microparticles during automatic mixing prior to use.
in tumor tissue.
Two monoclonal mouse antibodies - M‑4.184 and M‑3.170 - are used to Stability:
form the sandwich complex in the assay. unopened at 2‑8 °C up to the stated expiration date
Test principle after opening at 2‑8 °C 12 weeks
Sandwich principle. Total duration of assay: 18 minutes. on the analyzers 6 weeks
▪ 1st incubation: 10 µL of sample, a biotinylated monoclonal
ferritin‑specific antibody, and a monoclonal ferritin‑specific antibody Specimen collection and preparation
labeled with a ruthenium complexa) form a sandwich complex. Only the specimens listed below were tested and found acceptable.
▪ 2nd incubation: After addition of streptavidin-coated microparticles, the Serum collected using standard sampling tubes or tubes containing
complex becomes bound to the solid phase via interaction of biotin and separating gel.
streptavidin. Li‑heparin, Na‑heparin, K3‑EDTA and sodium citrate plasma.
When sodium citrate is used, the results must be corrected by + 10 %.
Criterion: Recovery within 90‑110 % of serum value or slope
0.9‑1.1 + intercept within < ± 2x analytical sensitivity (LDL) + coefficient of
correlation > 0.95.

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Ferritin
Ferritin

Serum, Li‑heparin, Na‑heparin and K3‑EDTA plasma is stable for 24 hours Bring the cooled reagents to approximately 20 °C and place on the reagent
at 20‑25 °C, 7 days at 2‑8 °C, 12 months at ‑20 °C. The samples may be disk (20 °C) of the analyzer. Avoid foam formation. The system
frozen twice. automatically regulates the temperature of the reagents and the
Sodium citrate plasma is stable for 7 days at 2‑8 °C, 12 months at ‑20 °C.8 opening/closing of the bottles.
The sample types listed were tested with a selection of sample collection Calibration
tubes that were commercially available at the time of testing, i.e. not all Traceability: The Ferritin assay (  03737551190) has been standardized
available tubes of all manufacturers were tested. Sample collection systems against the Ferritin assay (  11820982122). The Ferritin assay
from various manufacturers may contain differing materials which could (  11820982122) has been standardized against the Enzymun‑Test
affect the test results in some cases. When processing samples in primary Ferritin method. This in turn has been standardized against the
tubes (sample collection systems), follow the instructions of the tube 1st International Standard (IS) NIBSC (National Institute for Biological
manufacturer. Standards and Control) “Reagent for Ferritin (human liver)” 80/602.
Centrifuge samples containing precipitates before performing the assay. Recovery studies, including a published study,9 to assess traceability of the
Do not use heat‑inactivated samples. Elecsys Ferritin assay to more recent international standards
(2nd IS 80/578 and 3rd IS 94/572) have been conducted, with results
Do not use samples and controls stabilized with azide. showing very good agreement.
Ensure the samples, calibrators and controls are at 20‑25 °C prior to Every Elecsys reagent set has a barcoded label containing specific
measurement. information for calibration of the particular reagent lot. The predefined
Due to possible evaporation effects, samples, calibrators and controls on master curve is adapted to the analyzer using the relevant CalSet.
the analyzers should be analyzed/measured within 2 hours. Calibration frequency: Calibration must be performed once per reagent lot
Materials provided using fresh reagent (i.e. not more than 24 hours since the reagent kit was
See “Reagents – working solutions” section for reagents. registered on the analyzer). Renewed calibration is recommended as
follows:
Materials required (but not provided)
▪ after 8 weeks when using the same reagent lot
▪  03737586190, Ferritin CalSet, 4 x 1 mL
▪ after 7 days when using the same reagent kit on the analyzer
▪  11776452122, PreciControl Tumor Marker, for 2 x 3 mL each of
PreciControl Tumor Marker 1 and 2 or ▪ as required: e.g. quality control findings outside the defined limits
 05618860190, PreciControl Varia, for 2 x 3 mL each of PreciControl Quality control
Varia 1 and 2 For quality control, use PreciControl Tumor Marker or PreciControl Varia.
▪  11732277122, Diluent Universal, 2 x 16 mL sample diluent or In addition, other suitable control material can be used.
 03183971122, Diluent Universal, 2 x 36 mL sample diluent Controls for the various concentration ranges should be run individually at
▪ General laboratory equipment least once every 24 hours when the test is in use, once per reagent kit, and
following each calibration.
▪ Elecsys 2010, MODULAR ANALYTICS E170 or cobas e analyzer
The control intervals and limits should be adapted to each laboratory’s
Accessories for Elecsys 2010 and cobas e 411 analyzers: individual requirements. Values obtained should fall within the defined
▪  11662988122, ProCell, 6 x 380 mL system buffer limits. Each laboratory should establish corrective measures to be taken if
values fall outside the defined limits.
▪  11662970122, CleanCell, 6 x 380 mL measuring cell cleaning
solution Follow the applicable government regulations and local guidelines for
quality control.
▪  11930346122, Elecsys SysWash, 1 x 500 mL washwater additive
Calculation
▪  11933159001, Adapter for SysClean
The analyzer automatically calculates the analyte concentration of each
▪  11706802001, Elecsys 2010 AssayCup, 60 x 60 reaction vessels sample (either in µg/L or ng/mL).
▪  11706799001, Elecsys 2010 AssayTip, 30 x 120 pipette tips Limitations - interference
Accessories for MODULAR ANALYTICS E170, cobas e 601 and The assay is unaffected by icterus (bilirubin < 1112 µmol/L or < 65 mg/dL),
cobas e 602 analyzers: hemolysis (Hb < 0.31 mmol/L or < 0.5 g/dL), lipemia (Intralipid
▪  04880340190, ProCell M, 2 x 2 L system buffer < 3300 mg/dL) and biotin (< 205 nmol/L or < 50 ng/mL).
▪  04880293190, CleanCell M, 2 x 2 L measuring cell cleaning Criterion: Recovery within ± 10 % of initial value.
solution Samples should not be taken from patients receiving therapy with high
biotin doses (i.e. > 5 mg/day) until at least 8 hours following the last biotin
▪  03023141001, PC/CC‑Cups, 12 cups to prewarm ProCell M and administration.
CleanCell M before use
No interference was observed from rheumatoid factors up to a
▪  03005712190, ProbeWash M, 12 x 70 mL cleaning solution for run concentration of 2500 IU/mL.
finalization and rinsing during reagent change
There is no high-dose hook effect at ferritin concentrations up to
▪  12102137001, AssayTip/AssayCup Combimagazine M, 100000 µg/L (ng/mL).
48 magazines x 84 reaction vessels or pipette tips, waste bags
In vitro tests were performed on 19 commonly used pharmaceuticals. No
▪  03023150001, WasteLiner, waste bags interference with the assay was found.
▪  03027651001, SysClean Adapter M Iron2+‑ and iron3+‑ions at therapeutic concentrations do not interfere with the
Accessories for all analyzers: Elecsys Ferritin assay.
▪  11298500316, ISE Cleaning Solution/Elecsys SysClean, In rare cases, interference due to extremely high titers of antibodies to
5 x 100 mL system cleaning solution analyte‑specific antibodies, streptavidin or ruthenium can occur. These
effects are minimized by suitable test design.
Assay For diagnostic purposes, the results should always be assessed in
For optimum performance of the assay follow the directions given in this conjunction with the patient’s medical history, clinical examination and other
document for the analyzer concerned. Refer to the appropriate operator’s findings.
manual for analyzer‑specific assay instructions.
Resuspension of the microparticles takes place automatically prior to use.
Read in the test‑specific parameters via the reagent barcode. If in
exceptional cases the barcode cannot be read, enter the 15‑digit sequence
of numbers.

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Ferritin
Ferritin

Limits and ranges MODULAR ANALYTICS E170, cobas e 601 and cobas e 602 analyzers

Measuring range Repeatability Intermediate preci­
0.500‑2000 µg/L (ng/mL) (defined by the lower detection limit and the sion
maximum of the master curve). Values below the lower detection limit are
reported as < 0.500 µg/L (ng/mL). Values above the measuring range are Sample Mean SD CV SD CV
reported as > 2000 µg/L (ng/mL) (or up to 100000 µg/L (ng/mL) for 50‑fold µg/L µg/L % µg/L %
diluted samples). (ng/mL) (ng/mL) (ng/mL)
Lower limits of measurement Human serum 1 1.12 0.139 12.4 0.263 23.4
Lower detection limit of the test
Human serum 2 12.3 0.467 3.8 0.789 6.4
Lower detection limit: 0.50 µg/L (ng/mL)
The lower detection limit represents the lowest measurable analyte level Human serum 3 20.5 0.837 4.1 1.67 8.1
that can be distinguished from zero. It is calculated as the value lying two Human serum 4 392 8.14 2.1 16.9 4.3
standard deviations above that of the lowest standard (master calibrator,
standard 1 + 2 SD, repeatability study, n = 21). Human serum 5 1449 35.6 2.5 92.8 6.4
Dilution PreciControl Varia 1 140 2.31 1.7 3.53 2.5
Samples with ferritin concentrations above the measuring range can be PreciControl Varia 2 900 14.4 1.6 25.0 2.8
diluted with Diluent Universal. The recommended dilution is 1:50 (either
automatically by the MODULAR ANALYTICS E170, Elecsys 2010 or Method comparison
cobas e analyzers or manually). The concentration of the diluted sample A comparison of the Ferritin assay,  03737551190 (y) with the Ferritin
must be > 40 µg/L (ng/mL). assay,  11820982122 (x) using clinical samples gave the following
After manual dilution, multiply the result by the dilution factor. correlations:
After dilution by the analyzers, the MODULAR ANALYTICS E170, Number of samples measured: 134
Elecsys 2010 and cobas e software automatically takes the dilution into
account when calculating the sample concentration. Passing/Bablok11 Linear regression
Expected values y = 1.00x + 0.72 y = 0.99x + 4.11
Results of a study with the Enzymun‑Test Ferritin method on samples from τ = 0.984 r = 0.999
224 healthy test subjects (104 women - mainly premenopausal - and
120 men) are given below. The values correspond to the 5th and The sample concentrations were between approximately 2.68 and
95th percentiles.10 1891 µg/L (ng/mL).
Men, 20‑60 years: 30‑400 µg/L (ng/mL) Analytical specificity
Women, 17‑60 years: 13‑150 µg/L (ng/mL) Human liver ferritin: 100 % recovery
Each laboratory should investigate the transferability of the expected values Human spleen ferritin: 85 % recovery
to its own patient population and if necessary determine its own reference Human heart ferritin: 1 % recovery
ranges.
References
Specific performance data 1 Wick M, Pinggera W, Lehmann P. Ferritin in Iron Metabolism -
Representative performance data on the analyzers are given below. Diagnosis of Anemieas (second edition). Springer-Verlag 1995;ISBN
Results obtained in individual laboratories may differ. 3-211-82525-8 and ISBN 0-387-82525-8.
Precision 2 Arosio P, Levi S, Gabri E, et al. Heterogeneity of ferritin II:
Precision was determined using Elecsys reagents, pooled human sera and Immunological aspects. In: Albertini A, Arosio P, Chiancone E,
controls in a protocol (EP5‑A2) of the CLSI (Clinical and Laboratory Drysdale J (eds). Ferritins and isoferritins as biochemical markers.
Standards Institute): 2 runs per day in duplication each for 21 days (n = 84). Elsevier, Amsterdam 1984;33-47.
The following results were obtained: 3 Kaltwasser JP, Werner E. Serumferritin: Methodische und Klinische
Aspekte. Springer Verlag (1980).
Elecsys 2010 and cobas e 411 analyzers
4 Morikawa K, Oseko F, Morikawa S. A role for ferritin in hematopoiesis
Repeatability Intermediate preci­ and the immune system. Leuk-Lymphoma 1995;18(5-6):429-433.
sion 5 Borch-Iohnson B. Determination of Iron status: brief review of
Sample Mean SD CV SD CV physiological effects on iron measures. Analyst 1995;120(3):891-903.
µg/L µg/L % µg/L % 6 Cook JD, Skikne BS, Baynes RD. Iron deficiency: the global
(ng/mL) (ng/mL) (ng/mL) perspective. Adv-Exp-Med-Biol 1994;356:219-228.
Human serum 1 1.45 0.101 7.0 0.168 11.6 7 Albertini A, Arosio P, Chiancone E, et al. (eds). Functional aspects of
isoferritins. In: Jacobs A, Hodgetts J, Hoy TG. Ferritins and isoferritins
Human serum 2 11.9 0.411 3.5 0.798 6.7 as biochemical markers. Elsevier, Amsterdam 1984;113-127.
Human serum 3 19.2 0.780 4.1 1.47 7.7 8 Guder WG, Narayanan S, Wisser H, et al. List of Analytes;
Human serum 4 376 10.8 2.9 17.2 4.6 Preanalytical Variables. Brochure in: Samples: From the Patient to the
Laboratory. GIT-Verlag, Darmstadt 1996:14. ISBN 3-928865-22-6.
Human serum 5 1361 26.5 1.9 84.4 6.2
9 Blackmore S, Hamilton M, Lee A, et al. Automated immunoassay
PreciControl Varia 1 134 1.96 1.5 2.75 2.1 methods for ferritin: recovery studies to assess traceability to an
international standard. Clin Chem Lab Med 2008;46(10):1450-1457.
PreciControl Varia 2 858 15.1 1.8 21.7 2.5
10 Lotz J, Hafner G, Prellwitz W. Reference Study for Ferritin Assays.
Kurzmitteilung Clin Lab 1997;43(11):993-994.
11 Bablok W, Passing H, Bender R, et al. A general regression procedure
for method transformation. Application of linear regression procedures
for method comparison studies in clinical chemistry, Part III.
J Clin Chem Clin Biochem 1988 Nov;26(11):783-790.

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Ferritin
Ferritin

For further information, please refer to the appropriate operator’s manual for
the analyzer concerned, the respective application sheets, the product
information and the Method Sheets of all necessary components (if
available in your country).
A point (period/stop) is always used in this Method Sheet as the decimal
separator to mark the border between the integral and the fractional parts of
a decimal numeral. Separators for thousands are not used.
Symbols
Roche Diagnostics uses the following symbols and signs in addition to
those listed in the ISO 15223‑1 standard:
Contents of kit
Analyzers/Instruments on which reagents can be used
Reagent
Calibrator
Volume after reconstitution or mixing
GTIN Global Trade Item Number

COBAS, COBAS E, ELECSYS and PRECICONTROL are trademarks of Roche. INTRALIPID is a trademark of
Fresenius Kabi AB.
All other product names and trademarks are the property of their respective owners.
Additions, deletions or changes are indicated by a change bar in the margin.
© 2016, Roche Diagnostics

Roche Diagnostics GmbH, Sandhofer Strasse 116, D-68305 Mannheim

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