APTT LIQUID KIT

KIT FOR DETERMINATION

OF APTT (PARTICULATE
ACTIVATOR)
3. Centrifuge the specimen at 1000 rcf for 15 min.
Kit name Kit size Cat. No 4. Remove plasma from the tube within 60 min using a plastic
APTT LIQUID KIT 5 x 5 ml + CaCl2: 5x5ml 204-114 pipette and store in a plastic tube.
5 x 5.5 ml + CaCl2: 5x5.5ml 204-144V 5. Test plasma sample within 2 hours, otherwise store frozen and
thaw just prior to use.
INTRODUCTION Plasma pH will increase if exposed to air. Store samples stoppered.
From its origins through the work of Langdell and coworkers, and Do not delay mixing the blood with anticoagulant. Avoid foaming
later modified by others, the Activated Partial Thromboplastin Time the specimen. Use only plastic containers.
test (APTT, kaolin-kephalin time) has been widely used for a number
of years as a pre-surgical screen for assessing certain coagulation PROCEDURE
factors and in monitoring Heparin therapy. All Factors of the Kit is suitable for use with manual, mechanical or automated
Intrinsic Pathway are necessary for normal results when performing instrument for clot detection. See instrument manufacturers
the APTT test. instructions for full details.
The APTT test is also used to monitor Heparin therapy, showing
prolonged test results at approximately 0.1 units and above. Manual assay
The test is also used in the quantitative determination (Factor Assays) 1. Prewarm CALCIUM CHLORIDE 0,025M to 37°C. Mix gently.
of Factors VIII, IX, XI, XII and Prekallikrein. 2. Add 0.1 ml test plasma to cuvette and prewarm to 37°C.
3. Add 0.1 ml APTT-EA-REAGENT to the test plasma. Mix and
METHOD PRINCIPLE start timer.
APTT-P-REAGENT contains phospholipid and magnesium 4. Incubate the plasma-reagent mixture at 37°C for 5 minutes
aluminium silica as the activator for optimum sensitive Factor (activation time). For consistent results, test all plasmas with the
deficiencies and to heparin. The APTT-P test measures the clotting same activation time.
time of test plasma after the addition of APTT-P-REAGENT and 5. Forcibly add 0.1 ml prewarmed CALCIUM CHLORIDE
followed by the addition of calcium chloride. Deficiencies of 0,025M.
approximately 40% and lower of Factors VIII, IX, XI and XII will 6. Note time for clot formation.
result in a prolonged APTT. Heparin also results in a prolonged 7. Perform duplicate determinations.
APTT.
Calculation
REAGENTS Calculate the mean clotting time of duplicate determinations (to the
Package nearest 0.1 second) for each sample. A Normal Reference Range can
APTT LIQUID KIT also be reported for comparison. Do not report patient values relative
APTT-P-REAGENT 5 x 5 ml + CaCl2: 5x5ml to commercial control plasma clotting times. Controls are intended
APTT-P-REAGENT 5x 5.5ml + CaCl2: 5x5.5ml only for quality assurance of the test system, such as: temperature,
reagents, pipettes, instrument etc.
Working reagent preparation and stability
The reagents are ready to use. REFERENCE VALUES
The reagents when stored at 2-8 C are stable up to expiry date Method Mean Range (± SD)
printed on the package. Do not freeze. mechanical 30.3 26.1-34.5
Opened vials are stable for 30 days when stored at 2-8°C. photo-optical 33.3 28.5-38.1
Concentrations in the test These values should only be used as a guideline. Each laboratory
rabbit brain phospholipid 1.2 % should establish a Normal Reference Range (NRR) using
magnesium aluminium silica 0.03 % instrumentation, blood collection methods, and testing techniques
phenol 0.4 % used in that laboratory.
buffers 0.8 % A new NRR should be established with any change in
instrumentation, blood collection, techniques, anticoagulant and
Warnings and notes when changing to new lots of reagents.

Product for in vitro diagnostic use only.

The reagents must be used only for the purpose intended by QUALITY CONTROL
suitably qualified laboratory personnel, under appropriate CONTROL PLASMA NORMAL (Cat. No K-100) CONTROL
laboratory conditions. PLASMA-ABNORMAL LEVEL 1 and LEVEL 2 (Cat. No K-101

APTT-P-REAGENTS are designed to work at 37°. Frequently and K-102) should be tested in conjunction with patient plasmas. It is
check the temperature of all heating elements. recommended that controls be run at least each shift and a minimum

Mix gently before use. of once per 20 patient samples.

Always follow instrument manufacturer's instructions for proper Each laboratory should establish a control range for each control.
maintenance.
LIMITATIONS
ADDITIONAL EQUIPMENT A. Plasma samples with hematocrits outside the range of 20-55%

CALCIUM CHLORIDE reagent 0,025M (Cat. No K-355); may be improperly anticoagulated and should be adjusted

a manual, mechanical or photo-optical means of clot detection; appropriately.

timer; B. Turbid, icteric, lipemic, or hemolyzed specimens may generate

control plasmas; erroneous results.

general laboratory equipment C. Freezing and thawing plasma can affect results.
D. Acute inflammatory reactions can shorten APTT results because
SPECIMEN of elevated fibrinogen.
A. Anticoagulant- sodium citrate - 3.2% (0.105M). E. Sodium oxalate, EDTA, and heparin are not suitable
B. Specimen Collection: anticoagulants. Oral contraceptives, estrogen, pregnancy,
1. Obtain venous blood. coumarin type drugs, heparin, asparaginase, and naloxone have
2. Immediately mix 9 parts blood with 1 part anticoagulant, mix been reported to influence APTT results.
well by inversion of tube.

APTT-P page 1
PERFORMANCE CHARACTERISTICS LITERATURE
1. Heparin sensitivity: 1. Langdell, R., Wagner, R., Brinkhous, K.: Effect of Antihe-
For example, the following results were obtained on a photo-optical mophilic Factor on One Stage Clotting Tests. J. Lab. Clin. Med.
instrument with one lot of MTI reagent: 41: 637, 1953.
Heparin conc. (U/ml) APTT (s) 2. Proctor, R., Rapaport, S.: The Partial Thromboplastin Time with
0.0 31.9 Kaolin. Am. J. Clin. Path. 36: 212, 1961.
0.05 35.0 3. Ratnoff, O., Crum, S.: Activation of Hageman Factor by
0.1 39.9 Solutions of Ellagic Acid. J. Lab. Clin. Med. 63: 359, 1964.
0.2 53.2 4. Brandt J.T Triplet! D.A.: Laboratory Monitoring of Heparin.
0.3 68.5 Effect of Reagents and Instruments in the Activated Partial
0.4 85.3 Thromboplastin Time. AmerJ Clin Path 76:530, 1981.
0.5 106.5 5. Thompson, J.M.: The Control of Heparin Therapy by the
Each laboratory should establish its own heparin sensitivity ranges. Activated Partial Thromboplastin Time. Sensitivity of Various
Thromboplastins to Heparin. Standardization of Coagulation
Assays: An Overview. Edited by D.A. Triplett, College of
2. Factor sensitivity:
American Pathologists, Skokie, Ill. 1982, pp 195.
MTI reagents was evaluated on mildly and severely deficient
plasmas with the following results: 6. NCCLS: Collection, transport, and processing of blood
specimens for coagulation testing and performance of
Factor % activity APTT (s)
coagulation assays. 3rd edition. Approved guideline. NCCLS
VIII <1% 150
Document H21-A3. Wayne, PA, 1998.
VIII 20% 50.7 7. Young, D.S., Thomas, D.W., Friedman, R.B., et al: Effect of
IX <1% >150 Drugs on Clinical Laboratory Tests. Clin Chem 18:1041, 1972.
IX 20% 39.6 8. Banez, E.L., Triplett, D.A., Koepke, J.: Laboratory Monitoring
XI <1% 88.7 of Heparin Therapy The Effect of Different Salts of Heparin on
XI 20% 44.5 the Activated Partial Thromboplastin Time. AmerJ Clin Path
XII <1% >150 74:569,1980.
XII 20% 43.2 9. Wujastyk, J., Triplett, D.A.: Selecting Instrumentation and
Prekallikrein <1% 55.9 Reagents for the Coagulation Laboratory. Pathologist 37:398,
1983.
Furthermore, the sensitivity of APTT-P to Factor VIII has been 10. Christensen, R.L., Triplett D.A.: Factor Assay (VIII and IX)
determined as follows: Results in the College of American Pathologists Survey
% Factor VIII APTT (s) Program (1980-1982). AmerJ Clin Path 80 (Suppl): 633, 1983.
100% 34.3 11. NCCLS: One-Stage Prothrombin time (PT) Test and Activated
70% 37.8 Partial Thromboplastin Time (APTT) Test; Approved
50% 40.9 Guideline. NCCLS Document H47-A. NCCLS, Wayne, PA,
40% 43.4 1996.
30% 46.2
20% 50.7
10% 57.0 Date of issue: 01. 2008.
5% 63.6
1% 85.2
<1% >150
These values should only be used as guidelines. Each laboratory
should establish sensitivity to individual factors using instruments,
reagents, and techniques used in their laboratory.

WASTE MANAGEMENT
Please refer to local legal requirements.

MANUFACTURER

PZ MTI S.A.
ul. Wiosenna 22,
05-092 Łomianki, POLAND
tel.: +48 (0) 22 751 79 10
fax: +48 (0) 22 751 79 14
http://www.pzMTI.pl

01/08/01/08

APTT-P page 1