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Hemostat D-Dimer Centrifuge for 10 minutes at 3,000 x g or for 15 minutes at 1,500 x g. Re-
move plasma using a plastic pipette and store in a plastic tube. Plasma
Immunoturbidimetric Assay for the samples can be kept at 15…25°C for 8 hours. If specimen cannot be tested
within 8 hours, they should be stored frozen at -20°C or below.
Quantitative Determination of D-dimer
Test Procedure
Package Size HEMOSTAT D-DIMER can be used with semi-automated and with
[REF] 36002 2 x 1 ml Complete Kit automated coagulation analyzers.
[IVD] A) Applications for HumaClot Junior, HumaClot DuoPlus are displayed in the
Intended Use User Manual in combination with the calibration and pipetting scheme of
The HUMAN HEMOSTAT D-DIMER is an assay for quantitative this IFU (Instruction for Use)
determination of D-dimer in citrated plasma. It is intended for professional B) For HumaClot Junior (model HC1), HumaClot Duo Plus (model HC2),
use as an aid in diagnosis of thromboembolic events e.g. exclusion of DVT HumaClot Quattro (model HC4) and
(deep vein thrombosis) and PE (pulmonary embolism). It is suitable for C) the fully automated HumaClot Pro,
semi-automated and automated applications.
the application sheets can be accessed via www.human.de/aps-coag/
Principle
If the application sheets are not accessible via internet, they can be
HEMOSTAT D-DIMER consists of polystyrene particles coupled to mono- obtained free of charge from your local distributor. The application sheets
clonal antibodies specific to D-dimer. When a plasma containing D-dimer contain analyzer/assay-specific handling and performance information.
is mixed with the polystyrene particles and the reaction buffer, the particles Please also consult the user manual of the respective instrument.
will agglutinate, giving rise to increased light-scattering. When exposed to
a wavelength of 400 - 600 nm, the increase in turbidity is measured, which Calibration for the semi-automated instruments
is proportional to the amount of D-dimer in the sample. A) HumaClot Junior / HumaClot Duo Plus
Clinical Significance Concentration (ng/ml) DDU [CAL] [DIL]
Fibrin fragments containing D-dimer antigen are always present as a result Cal. 1 3,200* 200 µl 0 µl
of plasmin degradation of cross-linked fibrin. After injury, or when suffering Cal. 2 1,600* 200 µl 200 µl
from conditions associated with increased hemostatic activity, there is an
increase of D-dimer concentration. Elevated levels of D-dimer are found in Cal. 3 1 - -
clinical conditions such as deep vein thrombosis (DVT), pulmonary Measure Cal. 1 and Cal. 2 in duplicates, calculate mean and enter data
embolism (PE), disseminated intravascular coagulation (DIC) or trauma.1-4 into instrument. Cal. 3 is not measured, but the zero point is entered as
D-dimer levels increase during normal pregnancy but very high levels are 1 ng/ml. (HumaClot Junior: enter 1 mE; HumaClot DuoPlus: enter
associated with complications.7 D-dimer levels also increase with age.8 0.001 E).
A negative test result from a patient with a suspected thrombotic disorder * Use the LOT-specific value to specify the exact concentration in each dilution
has a high negative predictive value. 9 B) HumaClot Junior (model HC1) / HumaClot Duo Plus (model HC2) and
Contents HumaClot Quattro (model HC4)
[RGT] 2 x 1 ml D-dimer latex reagent (ready for use) Concentration (ng/ml) DDU [CAL] [DIL]
Polystyrene particles coated with monoclonal anti- Cal. 1 2,600 260 µl 40 µl
bodies (mouse)
Cal. 2 1,600 160 µl 140 µl
HEPES buffer 10 mmol/l
Bovine serum albumin, surfactant Cal. 3 800 80 µl 220 µl
Sodium azide 0.05% Cal. 4 400 40 µl 260 µl
[BUF] 2 x 2.5 ml Reaction buffer (ready for use) Cal. 5 220 22 µl 278 µl
HEPES buffer 100 mmol/l * Use the LOT-specific value to specify the exact concentration in each dilution
NaCl 400 mmol/l
A 5-point calibration with fixed calibrator points needs to be performed.
Sodium azide 0.05%
Because the original concentration of [CAL] is always higher than
[CAL] 1 x 1 ml Calibrator (lyophilised) 2600 ng/ml, the calibrator needs to be diluted with [DIL] accordingly for
Human plasma spiked with D-dimer every calibration step (cal. 1-5). The following formula can be applied:
HEPES buffer 33 mmol/l
Target concentration calibrator point
Read vial label for exact concentration!
Volume[CAL] (µl) =  x 300 µl
[DIL] 1 x 6 ml Diluent (ready for use) Lot specific [CAL] concentration
Phosphate buffer 20 mmol/l
Volume[DIL] (µl) = 300 µl - Volume[CAL] (µl)
Sodium azide <0.1%
Pipetting Scheme for the semi-automated instruments
Material required but not provided
A) HumaClot B) HumaClot Junior (model
[REF] 36012 HEMOSTAT D-DIMER CONTROL (4x1 ml) Junior, HC1), HumaClot Duo Plus
[HIGH] / [LOW]
HumaClot (model HC2), HumaClot
Photo-optical analyzer, application sheets: www.human.de/aps-coag/ DuoPlus Quattro (model HC4)
Reagent Preparation, Storage and Stability Pipette sample, 25 µl 50 µl
[RGT], [BUF] and [DIL] are ready-to-use. Prior to use, swirl vial gently to [CAL] or control
ensure homogenous suspension and allow [RGT] to equilibrate in the [HIGH] / [LOW]
instrument to working temperature. Reconstitute [CAL] with exactly 1 ml into the cuvette
distilled water (see section Limitations/Notes). Keep the vial at 15…25°C for Add [BUF] 100 µl 80 µl
15-30 minutes and gently swirl before use. Pre-warm 2 – max. 10 min. 150 sec.
Unopened reagents and calibrator are stable until the expiration date Transfer cuvette to measuring position and activate optics
when stored at 2…8°C.
Add pre-warmed 50 µl 40 µl
[RGT], [BUF] and [DIL] are stable for 2 weeks at 8…25°C or 4 weeks at 2…8°C [RGT]
after opening. Reconstituted [CAL] is stable for 12 hours at 4...25°C.
Mix well 15 x repeated Mixer is in cuvette
Avoid contamination of opened vials! pumping with
Specimen Collection pipette
Use 3.2% (0.109 M) buffered sodium citrate as anticoagulant. Obtain Results: E and ng/ml mE/min and ng/ml
venous blood by venipuncture. Immediately mix 9 parts blood with 1 part
Results
anticoagulant. Avoid foaming of the specimen. Please refer to CLSI
The results are reported in ng/ml D-dimer Units (DDU). Refer to the
Approved Guideline H21-A5 for more detailed instructions on specimen
instrument user manual and instrument-specific application sheet for
collection, handling and storage.
details. To convert results into FEU (Fibrinogen Equivalent Unit), multiply
the results DDU by 2.5.
Quality Control [CAL] provided in this kit contains material of human origin. It has been
It is recommended to use HEMOSTAT D-DIMER CONTROL HIGH/LOW at tested for HBsAg, HIV and HCV-antibodies and found to be negative using
regular intervals. Each laboratory should establish a control range to approved methods. However, the material should still be regarded as
determine allowable variation in the day to day performance of the test. potentially infectious.
Recalibration is suggested when control materials are assayed outside the References
acceptable range.
1. Heit J.A. et al.; Arch Path Lab Med. 123, 235-239 (1999)
Expected Values 2. Bounameaux H.J. et al., Thromb Haemostas 71, 1-6 (1994)
The normal level of D-dimer in the population is typically below 200 ng/ml
3. Pfitzner S.A. et al., Thromb Haemostas 78, 1069-1078 (1997)
DDU4,5 (equivalent to 500 ng/ml FEU). However, as there is no
internationally established standard for D-dimer, the concentration of 4. Lindahl T.L. et al., Scand J Clin Lab Invest 58, 307-316 (1998)
D-dimer may differ when determined using D-dimer assays from different 5. Gardiner C. et al., Brit J Haematol 128, 842-848 (2005)
manufacturers. Thus, each laboratory should establish its own reference
6. Meissner M.H. et al., J Trauma 54, 224-231 (2003)
intervals or cut-off levels.
7. Ballegeer V. et al., Thromb Haemostas 58, 1030-1032 (1987)
Performance Characteristics
Typical performance data of HEMOSTAT D-DIMER can be found in the 8. Kario K. et al., Thromb Res. 62, 501-508 (1991)
Verification Report, accessible via: 9. CLSI H59-A Quantitative D-dimer for the Exclusion of Venous
www.human.de/data/gb/vr/co-dd.pdf or Thromboembolic Disease Approved Guideline (March 2011)

www.human-de.com/data/gb/vr/co-dd.pdf
If the performance data are not accessible via internet, they can be
CO-DD INF 3600201 GB 03-2021-008 |
obtained free of charge from your local distributor.
Please refer to the respective application sheet for the HumaClot
Instruments for further information.
Example performance on HumaClot Pro: Sensitivity: 97%; specificity: 62%;
NPV: 99%; PPV: 41%.
Limitations / Notes
1. As there is no mixer in the cuvette for HumaClot Junior and HumaClot
DuoPlus, it is essential for reliable, precise manual results, to mix well
after addition of [RGT]. Mix 15 x using the pipette – avoid air bubbles!
2. D-dimer results should always be used with other clinical and
diagnostic information for forming a diagnosis and for patient
management.
3. Interferences: HEMOSTAT D-DIMER is insensitive to the following
substances: hemoglobin (up to 4 g/l), bilirubin (up to 0.1 g/l),
triglycerides (up to 2.5 g/l), low molecular weight heparin (up to
100 U/ml), and non-fractionated heparin (up to 100 U/ml). Turbid or
opalescent plasma may cause erratic results and should be interpreted
with caution; dilute the sample and re-assay.
4. Highly lipaemic plasma should be diluted with [DIL] and re-assayed
since elevated levels of triglycerides may cause depressed D-dimer
results.
5. The monoclonal antibody of HEMOSTAT D-DIMER has more than 100-
fold specificity for D-dimer (Fibrin or purified D-dimer) over Fibrinogen,
Fibrinogen D or Fragment E.
6. If double cuvettes are used for the HumaClot DuoPlus, do not touch the
cuvette during measurement. Use of single cuvettes ([REF] 18690) may
be preferred.
7. A new calibration curve should be established each time a new lot of
reagent kit is used or if the control plasmas are out of range. It is
recommended to re-calibrate at least every 3 months.
8. The distilled water used for reconstitution of [CAL] must be free from
preservatives and free from bacterial or other contamination.
Inadequate quality of water may lead to unreliable results and reduced
stability.
9. The presence of rheumatoid factor or HAMA (Human Anti-Mouse
Antibodies) may cause over-estimation of D-dimer test results.
10. All materials contaminated with human plasma should be inactivated
by validated procedures (i.e. autoclaving or chemical treatment)
according to applicable regulations.
Safety Notes
[RGT] [BUF] [CAL] [DIL]
P234 Keep only in original container.
P260 Do not breathe dust/fume/gas/mist/vapours/spray.
P262 Do not get in eyes, on skin, or on clothing.
P281 Use personal protective equipment as required.
P303+P361+P353 IF ON SKIN (or hair): Take off immediately all
contaminated clothing. Rinse skin with water/shower.
P305+P351+P338 IF IN EYES: Rinse cautiously with water for several
minutes. Remove contact lenses, if present and easy to do. Continue
rinsing.
P337+P313 If eye irritation persists: Get medical advice/attention.
P401 Store in accordance with local/regional/national/international
regulations.
P501 Dispose of contents/container in accordance with
local/regional/national/international regulations.

Human Gesellschaft für Biochemica und Diagnostica mbH