Fibrinogen

Presentation:
Ref.: 60121 R1 8 x 2 mL - R2 1 x 100 mL – R3 1 x 3.5 mL
FIBRINOGEN
Store: 2 - 8 º C. Clauss method
Diagnostic reagent for quantitative determination of Mixing, immediately, the blood with anticoagulant. Avoid foaming the QUALITY CONTROL
fibrinogen. specimen. Control sera are recommended to monitor the performance of assay
Centrifuge the sample at 3000 x g for 10 min and transfer the plasma to procedures:
Only for in vitro use in clinical laboratory (IVD) siliconized glass or plastic containers. CONTROL NORMAL REF: 60140
Turbid, icteric, lipemic or hemolyzed samples may generate erroneous CONTROL PATHOLOGICAL REF: 60150
TEST SUMMARY results. If control values are found outside the defined range, check the instrument,
Fibrinogen in presence of an excess of thrombin concentration, changes The sample is stable for 4 hours at room temperature (15-25º C) or 28 days reagents and technique for problems.
into Fibrin. if immediately frozen at below 20º C.
The time for clot formation in dilute plasma is inversely proportional to the Each laboratory should establish its own Quality Control scheme and
fibrinogen concentration in the sample. TEST PROCEDURE corrective actions if controls do not meet the acceptable tolerances.
The thrombin clotting time fibrinogen assay is based on the method The reagent can be used by manual procedure, mechanical, photo-optical
(Note 2)
originally described by Clauss.1 In the presence of high concentrations of or other means of end clot detection .
REFERENCE VALUES
thrombin, the time required for clot formation in dilute plasma is inversely 1. Dilute the citrated plasma and Control 1/10 with Imidazole buffer: 1
200 – 400 mg/dL . (2.0 – 4.0 g/L)
proportional to the fibrinogen concentration. 50 µL plasma + 450 µL Imidazole buffer. These values are for orientation purpose; each laboratory should establish
The diluted sample must be processed in 1 hour. its own reference range.
REAGENTS COMPOSITION 2. Prepare the following dilutions of the Calibrator in Imidazole buffer.
R1 Bovine thrombin ≈ 100 NIH u /mL Calibrator Dilution 1/40 1/30 1/20 1/10 1/5 CLINICAL SIGNIFICANCE
Fibrinogen (Factor I), protein synthesized by the liver, is the substance
IMIDAZOLE BUFFER (mL) 3.9 2.9 1.9 0.9 0.4 used in the blood to form a clot. Its determination is used to evaluate
R2 Imidazole Buffer
COAGULATION CAL (mL) 0.1 0.1 0.1 0.1 0.1 abnormal blood clotting.
Elevated Fibrinogen levels are observed in acute inflammations and in
R3 Caolin Solution Factor 10/40* 10/30* 10/20* 10/10* 10/5* pregnancy; low values are observed in trombolitic therapy, in hepatic
COAGULATION CAL REF: 60130 Concentration (mg/dL) 0.25* x c 0.33* x c 0.5* x c 1* x c 2* x c disease, in the congenital non fibrinogen, in DIC (Disseminated
Optional CONTROL N REF: 60140 Intravascular Coagulation) and in pancreatitis (low values)1.
(c = Calibrator value)
Clinical diagnosis should not be made on a single test result; it should
CONTROL P REF: 60150 3. Add 20 µL of R3 to 0.2 mL of each dilution, and allow to reach 37ºC
integrate clinical and other laboratory data.
for 4-6 minutes.
REAGENT PREPARATION AND STABILITY
4. Add 0.1 mL of R1 and time clot formation. Do not pre-warm thrombin PERFORMANCE CHARACTERISTICS
R1: Dissolve ( → ) the contents with 2.0 mL. of distilled water. Cap vial
R1. - Precision:
and mix gently to dissolve contents. Stability: 7 days at 2-8º C or 1 month at
Inter-assay (n=30)
–20º C, if immediately frozen and stored in the original container. Do not re- CALCULATIONS
1. Calculate the mean of duplicate clotting times immediately after Mean (U/L) 144 294 488
freeze.
R2: Mix before use. reaction. Use all five of the calibrator points to construct a log-log curve CV (%) 5.9 3.4 2.9
R3: Ready for use reagent. that plots fibrinogen concentration (mg/dL) vs. clotting time (s). - Accuracy: Results obtained using LABKIT reagents did not show
COAGULATION CAL (Calibrator): Dissolve ( → ) the contents with 1.0 mL 2. Draw the straight line of best fit. Examine the curve and, if necessary, systematic differences when compared with other commercial
omit non-linear points. The final curve must consist of at least three reagents.
of distilled water. Cap vial and mix gently to dissolve contents. Stability: 8 h
at 2-8º C. consecutives points. Constructing the curve with only the most linear INTERFERENCES
Signs of reagent deterioration: points will produce the best recovery on control and patient samples. Has been observed interferences in samples with fibrinogen degradation.
- Presence of particles and turbidity. 3. The following curve is only orientative. It will change with lot and Acute inflammatory reactions can elevate circulating fibrinogen. Hemolysis
- Quality control values outside established ranges. concentration of the calibrator, a well as, with the instrument used. can cause clotting factor activation and end point detection interference.
- Product colour variations. Time (s) Concentratation (mg/dL) Concentratation (g/L) High paraprotein levels,and drugs that activate the fibrinolytic system can
18.1 608 6.08 interfere with fibrinogen assays. A list of drugs and other interfering
All the components of the kit are stable until the expiration date on the 26.4 304 3.04 substances with the determination has been reported2,3.
label when stored tightly closed at 2-8º C, protected from light and 49.6 152 1.52
contamination prevented during their use. Do not use reagents over NOTES
the expiration date. 84.7 76 0.76
153 38 0.38 1. All labware must be clean and free of trace amounts of detergents.
2. Always follow instrument manufacturer’s instructions; the results must
ADDITIONAL EQUIPMENT 4. Find the clotting time of quality control and patient samples on the curve
be validated by the test laboratory.
- Coagulometer or stopwatch and bath at 37º C ± 0.5º C. and read the corresponding fibrinogen value.
5. If clotting times for the 1/10 dilution fall outside the linear curve, prepare BIBLIOGRAPHY
1/5 or 1/20 dilutions as needed. If the sample is diluted 1/5, divide the 1. Burtis A et al. Tietz Textbook of Clinical Chemistry, 3rd ed AACC 1999.
General laboratory equipment (Note 1)
result from the standard curve by 2; if the sample was diluted 1/20, 2. Young DS. Effects of drugs on Clinical Lab. Tests, 4th ed AACC Press, 1995.
multiply the curve result by 2 to get the final result. 3. Young DS. Effects of disease on Clinical Lab. Tests, 4th ed AACC 2001.
SPECIMEN
Plasma from venous puncture diluted 1/10 in trisodium citrate solution 3.8%
(105 mmol/L).

CHEMELEX, S.A.
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