Department: Clinical Laboratory and Blood Original Date :

Bank 2.10.2017
Revision Date :
SOP Code Number: DPP LB-HEM-008
14.3.2024
Effective Date :
Version : 5
14.4.2024
TITLE: APTT. (Activated partial
Review date :
thromboplastin time) on ( ACL TOP-350 ).
14.3.2026

1. Statement of PURPOSE
1.1 A high quality synthetic phospholipid reagent for the in vitro determination of Activated
Partial Thromboplastin Time (APTT) in human citrated plasma on IL Coagulation Systems.
The product is used for the evaluation of the intrinsic coagulation pathway,2 APTT substitution
test3 and the monitoring of heparin therapy.

2. APPLICABILITY
2.1 Laboratory technicians/specialists
2.2 Doctor In charge in Hematology

3. DEFINITION
3.1 The activated partial thromboplastin time (APTT), a global screening procedure used to
primarily to evaluate coagulation abnormalities in the intrinsic pathway, will also detect severe
deficiencies in factors II, V, VIII, IX, X, XI, XII or Fibrinogen.
The APTT also has been widely used to monitor the effectiveness of heparin therapy, where the
clotting time is prolonged in proportion to the level of heparin.
In patient receiving oral anticoagulants, the circulating levels of factors II, VII, IX and X are
depressed, therefore the APTT can be expected to be prolonged, the presence of non specific
inhibitors, such as the lupus like anticoagulant, may prolong the APTT depending on the nature of
the reagent. In summary the APTT is a clinically important screening test with wide applicability
for the diagnosis of coagulant disorders and therapeutic monitoring of both hemorrhagic and
thrombotic disease.

4. POLICIES
4.1 Assesment of APTT in citrated blood ( Na citrate 3.2 %,1: 9 )
4.2 Centrifugation 10 minutes at 2500g.
4.3 Plasma storage , stable for 8 hours at 2ºC - 8ºC and 4 hours at 15ºC - 25ºC.

DPP LB-HEM-008 (5) Page 1 of 7

Department: Clinical Laboratory and Blood Original Date :
Bank 2.10.2017
Revision Date :
SOP Code Number: DPP LB-HEM-008
14.3.2024
Effective Date :
Version : 5
14.4.2024
TITLE: APTT. (Activated partial
Review date :
thromboplastin time) on ( ACL TOP-350 ).
14.3.2026

5. PROCEDURES 5.1. principle

In the APTT test the incubation of plasma sample with an optimal
quantity of phospholipid, a negatively charged contact activator and
buffer initiates the activation of the intrinsic coagulation pathway.
After incubation at 37°C for a specific period of time calcium is then
added to trigger the coagulation process and the time required for clot
formation is measured.

DPP LB-HEM-008 (5) Page 2 of 7

Department: Clinical Laboratory and Blood Original Date :
Bank 2.10.2017
Revision Date :
SOP Code Number: DPP LB-HEM-008
14.3.2024
Effective Date :
Version : 5
14.4.2024
TITLE: APTT. (Activated partial
Review date :
thromboplastin time) on ( ACL TOP-350 ).
14.3.2026

5.2. CLINICAL SIGNIFICANCE

The activated partial thromboplastin time (APTT) is a general
coagulation screening test of the coagulation factors XII, XI, IX, VIII,
X, V, II and fibrinogen.
• A prolongation of the APTT is encountered in the following
situations:
– Congenital Deficiencies
◊ If the prothrombin time (PT) is normal, the following factors may be
deficient:
- factor VIII (STA® - Deficient VIII, REF 00725)
- factor IX (STA® - Deficient IX, REF 00724)
- factor XI (STA® - Deficient XI, REF 00723)
- factor XII (STA® - Deficient XII, REF 00722).
◊ If all these factors are normal, a deficiency in the following should be
considered:
- prekallikrein (Fletcher factor)
- HMW kininogen (Fitzgerald factor).
– Acquired Deficiencies and Abnormal Conditions
◊ Liver diseases
◊ Consumptive coagulopathy
◊ Fibrinolysis
◊ Circulating anticoagulants (LA type or circulating anticoagulant
against a factor)
◊ During heparin or oral anticoagulant therapy
◊ Treatments with thrombin inhibitors (e.g., hirudin, argatroban...).

DPP LB-HEM-008 (5) Page 3 of 7

Department: Clinical Laboratory and Blood Original Date :
Bank 2.10.2017
Revision Date :
SOP Code Number: DPP LB-HEM-008
14.3.2024
Effective Date :
Version : 5
14.4.2024
TITLE: APTT. (Activated partial
Review date :
thromboplastin time) on ( ACL TOP-350 ).
14.3.2026

6.3. Sample collection

Sample collection must be in conformity with the recommendations
for haemostasis tests.
• Collect blood (9 vol.) in 0.109 M (i.e., 3.2 %) trisodium citrate
anticoagulant (1 vol.). Use sample collection tubes made of plastic
or siliconized glass. [In the USA follow CLSI guideline documents
GP41-A6 (10) and H21-A5 (11)].
When monitoring heparin therapy, use preferably CTAD tubes,
specially designed sample collection tube to prevent heparin
inactivation (6).
• Centrifuge blood samples for 15 minutes at 2000-2500 g. Collect
plasma in plastic tubes.
• Plasmas remain stable for 4 hours at 20 ± 5 °C (9).
If on heparin therapy, plasmas remain stable for 2 hours at 20 ± 5 °C
when collected with citrate anticoagulant and for 4 hours at 20 ± 5
°C when collected with CTAD tubes.

6.4. Specimen Rejection:

6.4.1. Improperly filled specimen tubes, (Sodium citrate tubes
should be filled to75% or more of the fill line.)
6.4.2. Grossly hemolyzed specimens
6.4.3. Lipemic or icteric specimens should be evaluated on an
individual basis to determine acceptability.
6.4.4. Specimen contaminated with heparin or IV fluids.
6.4.5. Clotted specimens (check by gentle inversion).

DPP LB-HEM-008 (5) Page 4 of 7

Department: Clinical Laboratory and Blood Original Date :
Bank 2.10.2017
Revision Date :
SOP Code Number: DPP LB-HEM-008
14.3.2024
Effective Date :
Version : 5
14.4.2024
TITLE: APTT. (Activated partial
Review date :
thromboplastin time) on ( ACL TOP-350 ).
14.3.2026

4.5. Equipment & Supplies ( Stago C-Max 2)

Reagent/ Materials:
Calcium chloride solution (0.025M)
Activated PTT reagent.
Normal, high & low coagulation controls.
Coagulation Cuvettes.
Pipette and tips.
Kit Reagents
An Assay Value insert with a barcode is provided in the box. This
barcode contains the following information: lot number, kit code
number, reagent code number and expiration date.
STA® - PTT 5 : reagent containing cephalin (2) prepared from
rabbit cerebral tissues and a particulate activator (silica) in a
buffered medium, lyophilized.
5.6. Reagent Preparation
APTT Reagent: Each vial of APTT Reagent must be equilibrated at
15-25°C for at least 15 minutes and
mixed thoroughly before use.
Calcium Chloride: The reagent is ready for use.

5.7. Storage
Unopened reagents are stable until the expiration date shown on the
vial when stored at 2-8°C.
APTT Reagent: Opened reagent is stable 30 days at 2-8°C in the
original vial, 10 days at 15°C on the ACL TOP® Family
Do not freeze the reagent.

DPP LB-HEM-008 (5) Page 5 of 7

Department: Clinical Laboratory and Blood Original Date :
Bank 2.10.2017
Revision Date :
SOP Code Number: DPP LB-HEM-008
14.3.2024
Effective Date :
Version : 5
14.4.2024
TITLE: APTT. (Activated partial
Review date :
thromboplastin time) on ( ACL TOP-350 ).
14.3.2026

5.8.. Calibration
The pre-calibrated PTT values are identical for all the vials of each
lot. To enter the calibration data on the analyzer, scan the barcode
printed on the Assay Value insert across the instrument barcode
reader.

5.9. Calibration verification

The calibration data will be validated for the lot being used once the
two PTT control levels have been determined.
The calibration curve for PTT can be examined on the screen of the
analyzer in the “Calibration” menu
5.10. Quality control
Normal, low and high controls are recommended for a complete
quality control program. The laboratory establish its own mean
and standard deviation and establish a quality control program to
monitor laboratory testing. Controls should be analyzed at least once
every 8 hour shift in accordance with good laboratory practice.
5.11. Sample processing
Patients’ plasmas are tested undiluted. They are loaded in the
instrument. All samples tubes are barcoded. Firstly read the barcode
of the sample, the instrument through the LIS read the ordered
samples tests and process.
5.12. interpretation
5.12.1. Reference range.
A normal range study was performed using
SynthASil APTT reagent on ACL TOP
Family was 25.1 - 36.5 (seconds)
These results were obtained using a specific
lot of reagent. Due to many variables which
may affect clotting times, each laboratory
should establish its own normal range.

DPP LB-HEM-008 (5) Page 6 of 7

Department: Clinical Laboratory and Blood Original Date :
Bank 2.10.2017
Revision Date :
SOP Code Number: DPP LB-HEM-008
14.3.2024
Effective Date :
Version : 5
14.4.2024
TITLE: APTT. (Activated partial
Review date :
thromboplastin time) on ( ACL TOP-350 ).
14.3.2026

5.12.2. Therapeutic range

- If the patient is on heparin therapy a
baseline (pre therapy) level is
done .the therapeutic range for
heparin is 1.5-2 times this baseline
reading.
5.13.3. Panic results
PTT results more than 5o sec for outpatients
not on heparin therapy & more than 100 sec
for inpatients on heparin considered critical
and dealed with according to the panic result
policy.
5.13. Limitation When monitoring heparin therapy, any release
of platelet factor 4 (PF4) which is a potent
inhibitor of heparin, represents a major source
of error.
– Do not collect blood in glass, which might
cause this release; collect blood in plastic,
siliconized glass or CTAD tubes.
– Perform centrifugation within 1 hour after
sample collection if the blood was collected in
conventional citrate anticoagulant and within 4
hours if the blood was collected with CTAD
tubes.

6. RESPONSIBILITIES
6.1 Chief of Laboratory

8. REFERENCES
1. EBY C.: “Novel anticoagulants and laboratory testing”. International Journal of
Laborator Hematology, 35, 262-268, 2013.

DPP LB-HEM-008 (5) Page 7 of 7

Department: Clinical Laboratory and Blood Original Date :
Bank 2.10.2017
Revision Date :
SOP Code Number: DPP LB-HEM-008
14.3.2024
Effective Date :
Version : 5
14.4.2024
TITLE: APTT. (Activated partial
Review date :
thromboplastin time) on ( ACL TOP-350 ).
14.3.2026

2. CLSI Document EP05-A3: “Evaluation of precision performance of quantitative measuremen

methods; approved guideline”. Third Edition, 34, 13, 2014.
3. ADCOCK D.M., GOSSELIN R.: “Direct Oral Anticoagulants (DOACs) in the Laboratory: 2015
Review”. Thrombosis Research, 136, 7-12, 2015.

9. FORMS / EQUIPMENTS
9.1 ACL TOP-350 eqipment
9.2. PT and APTT reagents
9.3 Disposable cuvettes

10. REVISION
10.1 Will be reviewed after 2 years according to APP – 001

11. APPROVAL NAME & POSITION STAMP DATE

10.1.1 Dr. Nagah Mohammed
Head of Hematology dep.
10.1.2 Dr. Dr. Mohammed Ajlan
11.1. Prepared by
Asst. Lab Director for Med. Affairs
10.1.3 Hussin Nasser al Abayah
Clinical Lab. And B.B. Director

DPP LB-HEM-008 (5) Page 8 of 7

Department: Clinical Laboratory and Blood Original Date :
Bank 2.10.2017
Revision Date :
SOP Code Number: DPP LB-HEM-008
14.3.2024
Effective Date :
Version : 5
14.4.2024
TITLE: APTT. (Activated partial
Review date :
thromboplastin time) on ( ACL TOP-350 ).
14.3.2026

10.2.1 Mr. Ali Hamad Al Kulayb

11.2. Reviewed by
Quality & patient safety Director

10.3.1 Dr. Sayed Abdel Sabour Kinawy

Asst. Hospital Director for med. Service
11.3. Approved by
10.3.2 Dr. Mohammed Al Metlag
Hospital Director

DPP LB-HEM-008 (5) Page 9 of 7