Clinical Chemistry

2010/02/22 A93A01231BEN

A11A01900

ABX Pentra Ferritin 2 CP

Pentra C200

14 mL 10 mL

Intended Use: Diagnostic reagent for quantitative in vitro determination of Ferritin in serum and plasma by latex-enhanced immunoturbidimetric assay.

Clinical Interest (1)

Ferritin is an iron-containing protein with a molecular weight of approximately 450,000. It is found mainly in the human liver and spleen, where its function is to eliminate and store iron in the body, and is also found in small amounts in human serum. Consequently, the measurement of ferritin is considered to be useful in the diagnosis, treatment, assessment of disease progression, and postoperative prognosis for abnormality in iron metabolism such as iron deficiency anemia and hyperferremia as well as hepatitis and malignant tumors. ABX Pentra Ferritin 2 CP is a latex-enhanced immunoturbidimetric assay developed to accurately and reproducibly measure Ferritin levels in serum and plasma samples.

0.07 % w/v suspension of latex particles bound to anti-ferritin antibodies (rabbit) After measurements are taken, reagent cassettes should remain in Pentra C200 refrigerated tray. If used on other equipment, reagent cassettes should be capped and kept at 2-10C. Care should be taken not to interchange the caps of the reagent cassettes. Reagents with different lot numbers should not be interchanged or mixed. ABX Pentra Ferritin 2 CP should be used according to this reagent notice. The manufacturer cannot guarantee its performance if used otherwise.

Handling
1. Remove both caps of the cassette. 2. If present, remove foam by using a plastic pipette. 3. Place the cassette into the refrigerated Pentra C200 reagent compartment.

Method (2)
When an antigen-antibody reaction occurs between ferritin in a sample and anti-ferritin antibody which has been sensitized to latex particles, agglutination results. This agglutination is detected as an absorbance change, with the magnitude of the change being proportional to the quantity of ferritin in the sample. The actual concentration is then determined by interpolation from a calibration curve prepared from calibrators of known concentration.

Calibrator
For calibration, use: ABX Pentra Ferritin Cal, Ref. A11A01619 (not included) 4 x 1 mL

Reagents
ABX Pentra Ferritin 2 CP is ready-to-use. Reagent 1: Buffer solution Glycine buffer solution Reagent 2: Latex suspension

Control
For internal quality control, use: ABX Pentra Immuno II Control L/H, Ref. A11A01622 (not included) 1 x 3 mL (lyophilisate) + 1 x 3 mL (lyophilisate) Each control should be assayed daily and/or after a calibration.

Form-0846 Rev.4

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Clinical Chemistry

ABX Pentra Ferritin 2 CP

The frequency of controls and the confidence intervals should correspond to laboratory guidelines and countryspecific directives. The results must be within the range of the defined confidence limits. Each laboratory should establish a procedure to follow if the results exceed these confidence limits.

Packaging spoiling
In case of protective packaging spoiling, do not use the reagent if the damages might have an effect on the product performance.

Materials Required but not Provided

Automated clinical chemistry analyzer: Pentra C200 Calibrator: ABX Pentra Ferritin Cal, Ref.A11A01619 Control: ABX Pentra Immuno II Control L/H, Ref. A11A01622 NaCl solution: 9 g/L Standard laboratory equipment

Waste Management
Please refer to local legal requirements. This reagent contains less than 0.1 % of sodium azide as a preservative. Sodium azide may react with lead and copper to form explosive metal azides.

Specimen
Serum. Plasma in lithium heparin. Plasma in EDTA. The blood collection tube type has no influence on the test result. Stability of specimen (3): for up to 7 days at 20-25C. for up to 7 days at 2-8C. for up to 1 year at -20C. Repeated freezing and thawing should be avoided.

General Precautions
This reagent is for professional in vitro diagnostic use only. Diagnosis should only be made after taking clinical symptoms and the results of other tests into consideration. Specimens should be treated as potentially infectious (HIV, Hepatitis B virus, Hepatitis C virus, etc.) and handled with appropriate caution. The reagent cassettes are disposable and should be disposed of in accordance with the local legal requirements. Please refer to the MSDS associated with the reagent. Do not use the product if there is visible evidence of biological, chemical or physical deterioration.

Reference Range (4)

Because values could vary according to the age, the diet, the sex and the geographic repartition, each laboratory should establish its own reference ranges. The values given here are used as guidelines only. 10 - 120 ng/mL (g/L) Women: 20 - 250 ng/mL (g/L) Men: 6 months - 15 years: 7 - 140 ng/mL (g/L)

Performance on Pentra C200

The performance data listed below have been obtained on the Pentra C200 analyzer. Number of Tests: approximately 62 tests On Board Reagent Stability: Once opened, the reagent cassette placed in the refrigerated Pentra C200 compartment is stable for 31 days. Sample Volume: 18 L/test Minimum Interpretation Limit: The minimum interpretation limit (MIL) is evaluated using multiple determination of low concentration specimen and equals 3.5 ng/mL (g/L). Accuracy and Precision: Repeatability (within-run precision)

Storage and Stability

Reagents, in unopened cassettes, are stable up to the expiry date on the label if stored tightly closed between 2-10C, protected from light and contamination is avoided. Stability after opening: refer to the paragraph "Performance on Pentra C200".

Assay Procedure
Test instructions for other automated systems than Pentra C 200 are available on request (not available in the USA).

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Clinical Chemistry

ABX Pentra Ferritin 2 CP

6 specimens of low, medium and high concentration and 2 controls are tested 20 times according to the recommendations found in the Valtec protocol (5).
Mean value ng/mL (g/L) Control specimen 1 Control specimen 2 Specimen 1 Specimen 2 Specimen 3 Specimen 4 Specimen 5 Specimen 6 101.4 379.8 15.7 34.8 58.7 206.0 371.9 415.4 CV % 0.97 0.60 5.15 2.18 1.21 0.74 2.51 0.56

Interferences: Haemoglobin: No significant influence is observed up to 400 mol/L (690 mg/dL). Triglycerides: No significant influence is observed up to 7 mmol/L (612.5 mg/dL) (as Intralipid, representative of lipemia). Total Bilirubin: No significant influence is observed up to 500 mol/L (29.3 mg/dL). Direct Bilirubin: No significant influence is observed up to 500 mol/L (29.3 mg/dL). Other limitations are given by Young as a list of drugs and preanalytical variables known to affect this methodology (10, 11). Prozone Effect: No excess of antigen has been detected up to a concentration of 1036 ng/mL (g/L). Calibration Stability: The reagent is calibrated on Day 0. The calibration stability is checked by testing 3 control specimens. The calibration stability is 14 days. Note: A recalibration is recommended when reagent lots change, and when quality control results fall outside the range established. Application releasea : 01.xx

Reproducibility (total precision) 3 specimens of low, medium and high levels and 2 controls are tested 20 times according to the recommendations found in the CLSI (NCCLS), EP5-A2 protocol (6).
Mean value ng/mL (g/L) Control specimen 1 Control specimen 2 Specimen 1 Specimen 2 Specimen 3 102.07 380.45 58.64 212.70 419.34 CV % 1.30 2.12 2.57 1.92 2.06

Warning
It is the user's responsibility to verify that this document is applicable to the reagent used.

Measuring Range: The assay confirmed a measuring range from 3.5 to 500.0 ng/mL (g/L), with an automatic post-dilution up to 2500 ng/mL (g/L). The reagent linearity has been assessed up to 500 ng/mL (g/L) according to the recommendations found in the CLSI (NCCLS), EP6-A protocol (7). Correlation: 103 patient samples (serum) are correlated with a commercial reagent taken as reference according to the recommendations found in the CLSI (NCCLS), EP9-A2 protocol (8). Values ranged from 11.4 to 497.2 ng/mL (g/ L). The equation for the allometric line obtained using PassingBablock regression procedure (9) is: Y = 1.01 X - 0.91 (ng/mL (g/L)) with a correlation coefficient r2 = 0.9967.
a Modification

Reference
1. Thomas L. Ed. Clinical Laboratory Diagnostics. 1st ed. Frankfurt: TH-Books Verlagsgesellshaft, (1998): 278-281. Simo M, Joven J, Cliville X, Sans T, Automated latex agglutination immunoassay of serum ferritin with a centrifugal analyzer, Clin. Chem. (1994) 40: 625-629. Use of anticoagulants in diagnostic laboratory investigations. WHO publication WHO/DIL/LAB/99.1 Rev.2 (2002). Roberts WL, McMillin GA, Burtis CA, Bruns DE, Reference Information for the Clinical Laboratory, TIETZ Textbook of Clinical Chemistry and Molecular Diagnostics, 4th Ed. Burtis CA, Ashwood ER, Bruns DE, (Elsevier Saunders eds. St Louis, USA), (2006): 2269.

2. 3. 4.

from index A to B: Addition of minor index.

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Clinical Chemistry

ABX Pentra Ferritin 2 CP

5. Vassault A, Grafmeyer D, Naudin C et al. Protocole de validation de techniques (document B). Ann. Biol. Clin. (1986) 44: 686-745. 6. Evaluation of Precision Performance of Quantitative Measurement Method. Approved Guideline, CLSI (NCCLS) document EP5-A2 (2004) 24 (25). 7. Evaluation of the Linearity of Quantitative Analytical Methods. Approved Guideline, CLSI (NCCLS) document EP6-A (2003) 23 (16). 8. Method Comparison and Bias Estimation Using Patient Samples. Approved Guideline, 2nd ed., CLSI (NCCLS) document EP9-A2 (2002) 22 (19) . 9. Passing H, Bablock 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. 10. Young DS. Effects of Drugs on Clinical Laboratory Tests. 4th Edition, Washington, DC, AACC Press (1997) 3: 143-163. 11. Young DS. Effects of Preanalytical Variables on Clinical Laboratory Tests. 2nd Edition, Washington, DC, AACC Press (1997) 3: 120-132.

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