MEDT 20: HEMATOLOGY 2 LABORATORY

Activated Partial Thromboplastin Time (APTT)

Learning Outcomes

After the completion of this module, you will be able to:

1. Discuss the principle and clinical significance of activated partial
thromboplastin time (APTT)
2. Properly perform activated partial thromboplastin time (APTT)

Introduction

The activated partial thromboplastin time (APTT) is most useful procedure for routine
screening of coagulation disorders in the intrinsic system, for detecting the presence of
circulating anticoagulants (inhibitors), and for monitoring heparin therapy.
The normal range for the APTT may vary widely from one laboratory to another and
depends on the reagents used and clot detection method employed. The normal mean value
for the APTT will generally fall between 25 – 35 seconds (Brown), 20- 35 seconds
(Turgeon), 26 – 38 seconds (Rodaks).

Specimen

Fresh plasma from citrated whole blood is needed. Centrifuged unopened whole
blood specimens at 2,500 rpms for 20 minutes. Specimens should be tested within 2 hours
of collection

Materials, Regeants and Equipments

1. Water bath, 37˚ C

2. Calcium Chloride, 0.025 M
3. Partial Thromboplastin containing an activator
4. Test tubes
5. Stop watch

Principle

The calcium in whole blood is bound by the sodium citrate anticoagulant to prevent
coagulation. The plasma, after centrifugation contains all intrinsic coagulations factors
except calcium and platelets. Calcium, a phospholipid substitute for platelets (partial
thromboplastin) and an activator (to ensure maximal activation), are added to the plasma.
The time required for the plasma to clot is the activated partial thromboplastin time.

FNAA 2024
 MEDT 20: HEMATOLOGY 2 LABORATORY

Procedure

1. Centrifuge specimen as soon as possible after collection to obtain platelet poor

plasma.
2. Incubate a sufficient amount of 0.025 M of calcium chloride at 37 ˚ C.
3. Pipet 0.2 mL (200ul) of patient’s plasma into test tube.
4. Pipet 0.2 mL (200 ul) of partial thromboplastin (containing activator) into the test tube
containing the patient’s plasma.
5. Mix contents of the tube quickly and place in a 37 ˚ C water bath for 5 minutes.
6. After exactly 5 minutes, forcibly pipet 0.2 mL (200 ul) of prewarmed calcium chloride
into the tube and simultaneously start the stopwatch.
7. Mic the test tube immediately after adding calcium chloride. Allow the test tube to
remain in the water bath while gently tilting the tube every 5 seconds. At the end of
20 seconds, remove the test tube from the water bath. Quickly wipe off the outside of
the test tube with a clean gauze so that the contents of the tube can be clearly seen.
8. Gently tilt the test tube back and forth until clot forms, at which point the timing is
stopped.
9. Report the patients result in seconds.

Note: Same procedure goes with running the control.

Date
Name/s

FNAA 2024
 MEDT 20: HEMATOLOGY 2 LABORATORY

Year/Section/Cluster

1. Sources of error upon testing Activated Partial Thromboplastin Time (APTT) (3

points)

2. What is the clinical application or importance of testing APTT? (2 points)

3. Differentiate Partial Thromboplastin Time (PTT) from Activated Partial

Thromboplastin Time (APTT). (2 points)

4. Give some conditions that may lead to prolonged or an abnormal APTT result.

(3 points)

References:

 Brown, Barbara A. Hematology: Principles and procedures 6th Edition.

 Turgeon, M. (2012). Clinical hematology: Theory & procedures 5th Ed. Jones
& Bartlett Learning
 Elaine M. Keohane, Larry J. Smith, Jeanine M. Walenga 2016 Rodak's
Hematology Clinical Principles and Applications 5th Edition

FNAA 2024