Hemostat aPTT-EL Each laboratory should establish its own normal range using instrumentation,

blood collection methods, and testing techniques commonly used in that labo-
Manual and Automated Determination of ratory. The normal range should be re-established with any change in instru-
mentation, blood collection techniques, anticoagulant and should be at least
Activated Partial Thromboplastin Time using verified when changing to new lots of reagents.
3

Ellagic Acid Activator Limitations

Package Size Oral contraceptives, estrogen, pregnancy, coumarin type drugs, heparin, aspar-
2
[REF] 33002 6 x 4 ml for 240 Tests Complete Kit aginase, and naloxone have been reported to influence the aPTT test.
33012 6 x 4 ml for 240 Tests aPTT-EL Reagent Heparin Sensitivity
33013 6 x 10 ml for 600 Tests aPTT-EL Reagent The aPTT is commonly used to monitor heparin therapy since the prolongation
33022 4 x 30 ml CaCl2 of the aPTT is directly proportional to increasing amounts of heparin.
[IVD] In the presence of an adequate level of Antithrombin III the relative sensitivity of
a given aPTT reagent to heparin can be determined by establishing a heparin
Intended Use sensitivity curve. This is done by adding known amounts of heparin to pooled
The activated Partial Thromboplastin Time (aPTT) is a simple and versatile test normal plasma and performing an aPTT.
which is sensitive to deficiencies of all plasma clotting factors except Factor VII. Example: results obtained on a photo-optical instrument
However, it is mainly used to detect deficiencies in Factors VIII, IX, XI, XII, and
Prekallikrein (Fletcher Factor). Heparin (U/ml) HEMOSTAT aPTT-EL (sec.)
Test Principle 0.0 31.2
Activated partial thromboplastin time is performed by adding aPTT reagent 0.1 40.6
containing a plasma activator and phospholipid to the test specimen; the 0.2 56.2
phospholipid serves as a substitute for platelets. The mixture is incubated for
0.3 73.0
activation, then recalcified with calcium chloride and clot formation is timed.
0.4 85.9
aPTT-EL reagent can also be used to perform quantitative factor assays.
Each laboratory should determine its own heparin response curve using the
Contents
same heparin source used for therapy in that institution.1,2,5
[REF] 33002 33012 33013 33022
[RGT1] 6 x 4 ml 6 x 4 ml 6 x 10 ml Factor Sensitivity
[RGT2] 6 x 4 ml 4 x 30 ml Of the factors VIII, IX, XI, and XII, usually levels of 40% are sufficient to produce
normal aPTT results. If any of these factors is less than approximately 40%, the
[RGT1] aPTT-EL reagent
aPTT is prolonged.
rabbit brain phospholipid 0.007 %
ellagic acid 0.0037 % HEMOSTAT aPTT-EL was evaluated on mildly and severely deficient plasmas
buffers, salts and stabilisers with the following results:

[RGT2] CaCl2 0.02 mol/l Factor VIII IX XI XII Prekallikrein

salts and stabilisers % activity <1 <1 <1 <1 <1
Storage and Stability seconds 82.0 83.5 134.2 >200 69.5
Store at 2...8°C. Do not freeze. % activity 20% 20% 20% 20%
Opened vials are stable for 30 days when stored at 2...8°C. A yellow sediment seconds 44.8 40.9 47.8 36.2
may form after prolonged storage. Mix gently before use. Ready for use.
Quality Control
Specimen Collection: Use only plastic or siliconised glass containers and use Routine quality control is indispensable in coagulation testing.
3.2% (0.109 M) buffered sodium citrate as anticoagulant (sodium oxalate, EDTA
HEMOSTAT CONTROL PLASMA NORMAL (CPN) and ABNORMAL (CPA) should be
and heparin are not suitable).
tested in conjunction with patient plasmas. CPN is a normal plasma, whereas
Obtain venous blood by clean venipuncture. Immediately mix 9 parts blood with CPA is adjusted to mimic moderately deficient plasmas.
1 part anticoagulant. Avoid foaming the specimen.
Specimen Processing: Centrifuge for 15 minutes at 1500 g. Performance Characteristics
Typical performance data can be found in the Verification Report, accessible via
Remove plasma using a plastic pipette and store in a plastic tube. Cover speci-
mens to prevent pH changes that may affect test results. Turbid, icteric, lipemic, www.human.de/data/gb/vr/co-aptt.pdf
or haemolysed specimens may generate erroneous results. www.human-de.com/data/gb/vr/co-aptt.pdf
Specimens maintained at 22...24°C must be tested within 2 hours. For longer Automation
periods of time, plasma should be frozen at -20°C for up to 2 weeks or at -70°C
The test can be performed on mechanical and optical reading coagulation
for up to 6 months. Thaw samples rapidly at 37°C, swirl gently and test immedi-
systems. Please refer to the instructions given by the instrument manufacturer.
ately. Do not re-freeze.1,3
Applications for selected systems can be made available on request.
Test Procedure
References
Test samples and controls in duplicates.
1. Brandt J.T., Triplett, D.A., Amer. J. Clin. Path. 76, 530 (1981)
Pre- warm [RGT2] to 37°C.
2. Thomson J.M., CAP, Skokie, III.,195 (1982)
Pipette into pre-warmed test tube 3. NCCLS, NCCLS document H21-A3 (1998)
Plasma/ Control 0.1 ml 4. Young D.S. et.al, Clin. Chem. 18, 1041 (1972)
Incubate 1-2 min. at 37°C 5. Banez E.I. et al, Amer. J. Clin. Path. 74, 569 (1980)
Add [RGT1] 0.1 ml
Incubate 3-5 minutes* at 37°C CO-APTT INF 3300201 GB 06-2008-6 |
Add pre- warmed [RGT2] 0.1 ml
Start timer with addition of reagent. Record time required for clot formation.
*For consistent results, test all plasmas with the same time
Results
Calculate the mean time of duplicate aPTT determinations for each plasma and
report to the nearest 0.1 second.

Normal Range
When HEMOSTAT aPTT-EL was evaluated in a normal population, using both
mechanical and photo-optical clot detection systems, the following results were
obtained:
Mean (sec.) 2 S.D. Range
Mechanical 29.8 23.4-36.2
Photo-optical 30.7 26.1-36.3
Human Gesellschaft für Biochemica und Diagnostica mbH
Max-Planck-Ring 21 · 65205 Wiesbaden · Germany
Telefon +49 6122-9988-0 · Telefax +49 6122-9988-100 · e-Mail [email protected]