REF: K – 9560T

CRM diagnostic systems Ferritin

In vitro diagnostic reagents for the quantitative determination of human ferritin in Calibration. Quality control
serum by means of particle-enhanced turbidimetric immunoassay. Standardization: use CRM diagnostic systems Ferritin Calibrators. The method
Diagnostic Relevance was standardized against OMS 80/578 international standard.
Ferritin is a macromolecule with a molecular weight of at least 440 kD and is A multi-point calibration curve must be prepared using the Ferritin calibration
formed of apoferritin and an iron core of about 2500 Fe+3 ions set and newly opened reagents. This curve is stored in memory by the analyzer
It has been found a direct correlation between the plasma ferritin concentration and recalled for later use. Calibration curves are stable for up to 14 days, after
and the quantity of available iron stored in the body so that its determination is which a new curve must be generated. Additionally, recalibration must be
used for diagnosis and monitoring of iron deficiency and iron overload. performed whenever reagent lots are changed or quality control so indicates.
Additional parameters (transferrin, transferrin saturation, and hematological For quality control use CRM diagnostic systems Ferritin Controls or other
investigations) could be required for the diagnosis of disturbances of distribution. suitable control material. The control intervals and limits must be adapted to the
In a comparison of the various parameters available for the determination of the individual laboratory requirements. Values obtained should fall within established
body’s iron stores, plasma ferritin was the most efficient parameter, limits. Each laboratory should establish corrective measures to be taken if
demonstrating a sensitivity of 80 %, and a specificity of 96 %. values fall outside the limits. Control must be assayed and evaluated as for
The serum concentrations of ferritin are found to be elevated in patients with patient samples.
infections, inflammation or in hepatic or chronic renal diseases. The
determination of ferritin is particularly useful in the diagnosis of iron therapy, for
Calculation
the determination of iron reserves in high-risk groups, and in the differential The turbidimetric analysers automatically calculate the Ferritin concentration of
diagnosis of anaemia. each sample.

Principle Reference Intervals

The Ferritin test is based upon the reactions between Ferritin in the sample and The determination of reference ranges for ferritin concentrations of clinically
latex-covalently bound antibodies against human Ferritin. Ferritin values are healthy individuals is very difficult. Ferritin concentrations are age- and sex-
determined turbidimetrically using fixed-time measurement with sample blank dependent and exhibit a wide range of distribution.
correction. The relationship between absorbance and concentration permits a Children: Cord blood contains 100 a 250 ng/mL. In the first two months of life
multipoint calibration with a measuring range between 0 and 500 ng/mL. The there is a rise of up to 600 µg/L, followed by a fall of down to 1 µg/L (Hb-
measuring temperature is 37ºC. The assay can be performed on different neosynthesis).
instruments allowing turbidimetric measurements at 500 to 600 nm. Children and adolescents 15 - 120 ng/mL.
(6 weeks to 18 years of age)
Reagents Men 30 - 300 ng/mL
Each Ferritin kit contains: Women (Pre-menopausal) 10 - 160 ng/mL
A.- Buffer - 25 mL of phosphate buffer, pH: 6.7, containing protein stabilizers Women (Post-menopausal) 30 - 300 ng/mL
and 0,09 % sodium azide as preservative. These data are to be interpreted as a guide. Each laboratory should establish its
B.- Latex reagent – 7,5 mL of a suspension of latex microparticules covalently own reference intervals.
bound anti-ferritin antibodies suspended in a neutral aqueous solution, with 0,09
% sodium azide as preservative.
Assay range and Sensitivity
The assay range is established from 0 to 500 ng/mL.
Precautions Calculating the mean plus 2SD of twenty replicates of zero standard resulted in
For in vitro diagnostic use only. Do not pippete by mouth. Reagents containing a lower limit of detection of 5,2 ng/mL.
sodium azide must be handled with precaution. Sodium azide can form
explosive azides with lead and copper plumbing. Since absence of infectious
Specificity
agents cannot be proven, all specimens and reagents obtained from human The assay is specific for Ferritin determination. Interference from billirrubin,
blood should always be handled with precaution using established good hemolisis or rheumatoid factors has not been observed.
laboratory practices. Disposal of all waste material should be in accordance with Other substances can interfere. For a comprehensive review of interfering
local guidelines. substances, refer to the publication by Young.
As with other diagnostic tests, results should be interpreted considering all other Assay Precision
test results and the clinical situation of the patient. Intra-assay coefficients of variation (CV) for three samples (ferritin values
Materials required but not provided ranging from 30 to 300 ng/mL) were between 1,5 and 7,5 %; daily calibrated
Automatic analyzer. inter-assay CVs were between 3,2 and 8,5 %.
Saline solution. Assay Linearity
Set of calibration. Linearity was evaluated using serial dilutions, prepared with saline solution, of
Set of controls. two different samples, which contained values of Ferritin in the range of analysis.
Storage and Stability Linear regression values of Ferritin ng/mL vs concentration yielded correlation
The ferritin reagents should be stored tightly capped at +2..+8ºC when not in coefficients, r>0,999, for all samples. Within the assay’s measuring range, the
use. Do not freeze. Reagents in the original vials are stable to the expiration deviations of measurement from theoretical values did not exceed the 10 %
date on the vial label when capped and stored at +2...+8ºC. Immediately level. In addition, the system did not show prozone phenomenon at least up to
following the completion of an assay run, the reagent vials should be capped 5500 ng/mL.
until next use in order to maximize curve stability. Once opened the reagent can Method comparison
be used within 1 month if stored tightly closed at +2...+8ºC after use. 46 samples (conc. aprox. 5 to 500 ng/mL) were correlated with a nephelometric
The ferritin buffer reagent should be clear and colourless. Any turbidity may be commercial procedure. When comparing the results by lineal regression the
sign of deterioration and reagent should be discarted. results was: y = 1,01x + 0,34 and r=0.995.
The ferritin latex reagent should have a white, turbid appearance free of granular
particulate. Visible agglutination or precipitation may be a sign of deterioration, Analytical characteristics have been obtained in a single experiment in a
and the reagent should be discarted. Cobas-Mira plus analyser. As is well known the analytical characteristics
Specimens of a clinical chemistry reagent depend on both the reagents and the
Specimens should be collected by venipuncture following good laboratory instrument used. Multicenter studies indicate important differences in
practices. Suitable assay specimens are human serum samples, as fresh as analytical characteristics among similar instruments. Therefore, the data
possible (stored up to 7 days at +2...+8ºC) or deep-frozen. Any additional expressed in the present document should be interpreted as a guide
clotting or precipitation, which occurs due to the freeze/thaw cycle, should be example.
removed by centrifugation prior to assay.
Very lipemic specimens, or turbid frozen specimens after thawing, must be Literature
clarified before the assay by high-speed centrifugation (15 min at approx. 15.000 Wick M, Pinnggera W, Lehmann P. Ferritin in iron metabolism. Diagnosis of anemias. 2nd ed.
rpm). Springer-Verlag. Wien 1994.
Miles LEM, et al. Measurement of serum ferritin by a 2-site immunoradiometric assay. Anal
Procedure Biochem 1974; 61:209-224
Milmann N, Sondergaard M, Sorensen CM. Iron stores in female blood donors evaluated by
The reagents are ready to use as supplied. Reagents should be gently shaken serum ferritin. Blut 1985;51:337-345.
before each use. Young DS. Effects of Drugs on Clinical Laboratory Test. 5th Edition, AACC Press, 2000.
Follow the instructions of the operator’s manual to load the cartridge, technique Sonderdruck aus DG Klinische Chemie Mitteilungen 1995; 26: 207 – 224.
programation, calibration, sample measurement and control.
Significados de los símbolos indicados en las etiquetas. Explanation of symbols used on labelling. Explication des symbols figurant sur les etiquettes. Spiegazione dei simboli utilizzati sull’eticheta. Significado dos símbolos indicados nas etiquetas. Erläuterung der symbole auf den etiketten.

LOT IVD REF REAG CAL Buffer LYOPH Conc. Control H / Control L
Fecha de Caducidad Temperatura de almacén Número de Lote Para Diagnóstico In Vitro Número de catálogo Conformidad Europea Fabricado por Reactivo Calibrador Tampón Liofilizado Concentración Control Alto / Control Bajo
Expirate Date Storage Temperature Lot Number For In Vitro Diagnostic Catalog Number European Conformity Manufactured by Reagent Calibrator Buffer Lyophilised Concentration Control High / Control Low
Date de Péremption Temperature de Conservation Número de Lot Usage In Vitro Numéro de catalogue Conformité aux normes européennes Fabriqué par Réactif Calibrateur Tampon Lyophilisé Concentration Contrôle élevè / Contrôle Bas
Data di Scadenza Temperatura de Conservazione Numero di Lotto Per Uso Diagnostico In Vitro Numero di catalogo Conformità europea Fabbricato da Reagenti Calibradore Tampone Liofilo Concentrazione Controllo Alto / Controllo Basso
Data Expiração Temperatura de Conservação Número de Lote Utilizar em Diagnostico In Vitro Número de catálogo Comformidade com as normas europeias Fabricado por Reagente Calibrador Buffer Liofilizado Concentraçao Controlo Alto / Controlo Baixo
Verwendbar bis Lagertemperatur Chargen-Nr In Vitro Diagnosticum Katalognummer CE-Konformitätskennzeichnung Hergestellt Reagenz Kalibrator Puffer Lyophilisiert Koncentration Kontrolle Hoch / Kontrolle Niegrid

BL, S.L. C/ Calahorra 4-26006 Lo-Spain 2010/09

 Ferritin / Ferritina
Procedure for / Procedimiento para
COBAS MIRA PLUS

GENERAL CALCULATION (continued)

Measurement mode: Absorb Antigen excess NO

Reaction mode: R-S-SR1 Conversion factor: 1.00000
Calibration mode: LOGIT/LOG5 Offset: 0.00000
Reagent blank: REAG/DIL Normal range low: NO

Cleaner: NO Normal range high: NO

Wavelenght: 600 nm Number of steps: 1
Decimal position: 2 Calc. Step A: KINETIC

Units: ng/ml Reading first: 3

Reading last: 12

ANALYSIS Reaction limit: 3.5000 ∆A

Post. Dil. Factor: 10.00 Point: T2

Conc. factor: NO CALIBRATION

SAMPLE Vol: 25 µL Calibr. Interval: ON REQUEST

Cycle: 3 Reagent blank:

DILUENT Name: Saline Reag. Range low: NO High: NO

Vol: 20 µL Blank range low: NO High: NO

REAGENT Vol: 200 µL

Cycle: 1 STD POS**:

START REAGENT Vol: 75 µL 1: 0.0*** 2: 37.5***
Cycle: 1 3: 75.0*** 4: 150.0***

DILUENT Name: Saline 5: 300.0*** 6: 500.0***

Vol: 0.0 µL 7: NO 8: NO
CALCULATION

Sample limit: NO REPLICATE: SINGLE

Reaction Direction: INCREASE Deviation:
Check: ON Correction std:

(**) Select one position on the standard rack

(***) Standard values on the labels / inserts

Revision 2010/09