APTT

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APTT

CONTENTS
APTT
Activated Partial Thromboplastin Time
REF 3510201 APTT 10 x 4 mL
(APTT)
Ellagic acid
For in vitro diagnostic use only

PRINCIPLE 3. Plasma Storage


- Plasma samples may be stored at room temperature for up to 2
The capacity of blood to form a fibrin clot by means of the intrinsic hours; at 2-8ºC for up to 4 hours; frozen at ≤ -20 °C for up to 2
hemostatic pathway requires coagulation factors I, II, V, VIII, IX, X, months. Plasma may be re-centrifuged prior to freezing to assure
XI and XII, platelet lipids and calcium4. The assay is performed by that all cells are removed. Quick thaw frozen samples and test
the addition in the sample of a suspension of rabbit brain cephalin immediately.
with a surface activator1. - Avoid contact of samples with glassware.
The APTT has proven to be a simple and highly reliable - Samples should not stand at +37 °C for more than five minutes.
measurement of the intrinsic coagulation mechanism5.
INTERFERENCES
REAGENT COMPOSITION
APTT Rabbit brain cephalin and ellagic acid activator with  Delay in testing, difficulty in specimen collection, or venipucture
above the site of a heparin lock may result in falsely prolonged
buffer. Stabilizers and preservatives.
APTT results7.
Optative. Calcium Chloride 0.02 mol/L Ref 3510401  The APTT may also be influenced by certain drugs and
Plasma Control Level 1 Ref. 3520101, medications8. APTT results can vary with anticoagulation
Plasma Control Level 2 Ref. 3520201. therapy depending upon the type and dosage of anticoagulant,
the route of administration and the time of administration of the
STORAGE AND STABILITY last dose.
Store at 2-8ºC. The reagent is stable until the expiry date
stated on the label. ADDITIONAL EQUIPMENT
The reconstituted APTT reagent is stable for 5 days when stored in
 Coagulometer or stopwatch and bath at 37ºC ± 0.5ºC.
the original container at 2-8ºC. Do not freeze.
 General laboratory equipment.
REAGENT PREPARATION PROCEDURE
1. Reconstitute the contents of APTT with 4,0 mL of distilled water. This procedure pertains to manual or semi-automated coagulation
2. Replace the stopper and thoroughly mix the vial contents. Let systems. For best results, duplicate samples are recommended.
stand for no less than 30 minutes prior to use to assure
complete hydration of the contents. 1. Pre-incubate the Calcium Chloride 0.02 mol/L to 37oC for at
least 10 minutes.
SAMPLES 2. Pipette 50 L of sample or control plasma into a test cuvette.
Incubate at 37oC for 3 minutes.
Test plasma should be prepared from citrated whole blood. Avoid 3. Add 50 L of APTT reagent to the cuvette containing the
use heparin, EDTA or oxalate. Mix nine parts of freshly collected plasma.
patient blood with one part of 3.2% trisodium citrate. 4. Incubate the mixture at 37oC for 3 minutes.
Refer to National Committee for Clinical Laboratory Standards 5. Rapidly add 50 L of the pre-incubated Calcium Chloride and
(NCCLS) guidelines H3-A3 and H21-A3. simultaneously start the timer.
6. Record the clotting time in seconds.
1. Blood Collection
- Draw venous blood into a plastic or siliconized syringe. For semi-authomatic system refer to your instrument manual.
- Immediately transfer the blood into a tube containing sodium
citrate solution. or CALCULATIONS
- Draw venous blood into a commercial vacuum tube containing Calculate the mean clotting time of duplicate samples and controls.
sodium citrate solution. Differences between duplicate results should be less than 5%.
- Mix well by inversion Repeat the test if necessary.
Insure that a full draw has been obtained since the ratio of 9 parts
blood to 1 part citrate is critical. The APTT result may be reported as two different ways:
1. Seconds, observed clotting time.
2. Plasma Preparation
- Centrifuge the blood specimen at 2,500 x g for no less than 15 2. APTT ratio:
minutes at room temperature as soon as possible after APTT of the patient in sec
APTT ratio 
collection. Store in an unopened tube at room temperature. APTT of normal plasma in sec
- If immediate testing is to be done, the plasma may remain on the
packed cells. Otherwise plasma should be separated from the
cells. To separate the plasma, use a plastic transfer pipette and
remove plasma to a plastic tube.

QUALITY SYSTEM CERTIFIED LINEAR CHEMICALS, S.L.U. Joaquim Costa 18 2ª planta. 08390 Montgat (Barcelona) SPAIN
ISO 9001 ISO 13485 Telf. (+34) 934 694 990; E-mail: [email protected] ; website: www.linear.es NIF-VAT:B60485687
REFERENCE VALUES
APTT results are influenced by the method of clot detection and - Traceability: Stago PTTA (micronised silica activator)
can vary from laboratory to laboratory. In general an APTT test
performed on a photo-optical coagulometer will give clotting time for - Accuracy
normal plasma in the range of 25 to 43 seconds. Therapeutic Results obtained with this reagent did not show significative
ranges for monitoring oral anticoagulation therapy will vary from differences when compared with reference reagents. Details of
laboratory to laboratory, therefore it is essential that each laboratory the comparison experiments are available on request.
establish relevant APTT ranges for its respective patient population.
Abnormal results obtained with a plasma from a patient not on REFERENCES
anticoagulant therapy may indicate a factor deficiency or the 1. Human Blood Coagulation, Hemostasis and Thrombosis, 3rd ed.
presence on an inhibitor. The result may also be due to the effects R Biggs, CR Rizza, Editors, Blackwell Scientific Publications,
of certain drugs and medications. Additional procedures such as London (1984).
the PT test and mixing studies using factor deficient plasma are 2. Cole E, Hall ER, Wu KK, Principles of Antithromboltic Therapy. In
usually required. Wu KK, Thomboembolic Disorders, PSG Publishing Co. Inc.,
Litteton, p 91 (1984).
QUALITY CONTROL 3. Triplett DA, Heparin: Clinical use and Laboratory Monitoring. In
Triplett DA, Laboratory Evaluation of Coagulation, American
Control sera are recommended to monitor the performance of Society of Clinical Pathologists Press, Chicago p 272 (1982).
assay procedures. They should be used as sample. 4. Hougie C, The Biochemistry of Blood Coagulation, In Laboratory
Evaluation of Coagulation, American Society of Clinical
REF 3520101 PLASMA CONTROL LEVEL 1
Pathologists Press, Chicago p 2 (1982).
REF 3520201 PLASMA CONTROL LEVEL 2 5. Owen CA, Bowie EJW, Thomson JH, The Diagnosis of Bleeding
Disorders, Little Brown and Company, Boston p 110 (1975).
6. Harker LA, Hemostasis Manual, FA Davis Co, Philadelphia p 62
Each laboratory should establish its own Quality Control scheme
(1974).
and corrective actions if controls do not meet the acceptable
7. Triplett DA, Harms CS, Procedures for the Coagulation
tolerances. Laboratory, American Society of Clinical Pathologists Press,
Chicago, p 7 (1981).
CLINICAL SIGNIFICANCE 8. Young DS, Pestaner LC, Gibberman V, Effects of Drugs on
Clinical Laboratory Tests, Clin Chem 21; 1D (1975).
The APTT reagent is an in vitro diagnostic assay intended for use in
determining activated partial thromboplastin time (APTT) and
coagulation factor assays that are based on a modified APTT.
The activated partial thromboplastin time (APTT) is used as a
general screening test for the detection of coagulation abnormalities
in the intrinsic pathway. The APTT is sensitive to deficiencies or
abnormalities of factors VIII, IX, XI, XII, X, and II, prekallikrein, high
molecular weight kininogen (HMWK), and fibrinogen. APTT is also
sensitive to inhibitors of blood coagulation such as lupus inhibitor
and fibrin/fibrinogen degradation products (1). The APTT is the most
widely used method for monitoring intravenous heparin
anticoagulation therapy2, 3.

ANALYTICAL PERFORMANCE
- Linearity
 0.2-0.6 IU Unfractionated heparin
 0.01-0.65 IU Factor VIII

- Analytical sensitivity
 < 0. 01 IU Factor VIII

- Normal APTT time Mean


 30-44 sec.

- Heparin sensitivity:
The anticoagulant action of heparin depends on many factors. Each
laboratory should determine the relative heparin sensitivity by
adding known amounts of unfractionated heparin to pooled normal
plasma and determining the elevation in clotting time using the
APTT reagent.

Heparin Concentration APTT – Clotting Time


(units / mL) (in seconds)
0.4 IU 68-75

C35102-3/1202
R1.ing

QUALITY SYSTEM CERTIFIED LINEAR CHEMICALS, S.L.U. Joaquim Costa 18 2ª planta. 08390 Montgat (Barcelona) SPAIN
ISO 9001 ISO 13485 Telf. (+34) 934 694 990; E-mail: [email protected] ; website: www.linear.es NIF-VAT:B60485687

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