200 Tests

APTT ‘New’ 12602003

Intended Use Procedure

The APTT reagent is an in vitro diagnostic assay intended for use in determining This procedure pertains to the Mispa Clog Plus coagulation system:
activated partial thromboplastin time (APTT) on the Mispa Clog Plus Analyzer.
1. Pre-incubate the Calcium Chloride (0.02M) to 37 OC for at least 10 minutes.
Summary
2. Pipette 100 µL of sample or control plasma into a coagulation microcuvette
The activated partial thromboplastin time (APTT) is used as a general screening
and pre-incubate in channel from 1 to 4 at 37 OC for 1 to 2 minutes.
test for the detection of coagulation abnormalities in the intrinsic pathway.
3. Add 100 µL of the APTT reagent to the microcuvette containing the plasma.
The APTT is sensitive to deficiencies or abnormalities of factors VIII, IX, XI,
Maintain the suspension of the reagent by magnetic stirring or mixing by
XII, X, and II, prekal likrein, high molecular weight kininogen (HMWK), and
inversion immediately prior to use.
fibrinogen. APTT is also sensitive to inhibitors of blood coagulation such as
lupus inhibitor and fibrin/fibrinogen degradation products (1). 4. Incubate the mixture at 37 OC for 3 minutes.
5. Add a magnetic stirrer to the microcuvette.
Principle
6. Transfer the microcuvette into the reading channel (Filter A).
The APTT is the most widely used method for monitoring intravenous heparin 7. When requested on the display, rapid ly add 100 µL of the preincubated
anticoagulation therapy (2, 3).The capacity of blood to form a fibrin clot by
Calcium Chloride (0.02M).
way of the intrinsic hemostatic pathway requires coagulation factors I, II, V,
8. Record the clotting time which wil l appear automatical ly on the display
VIII, IX, X, XI and XII, platelet lipids and calcium(4). The assay is performed by
the addition of a suspension of rabbit brain cephalin with a surface activator(1). once the clot is formed.
The APTT
has proven to be a simple and highly reliable measurement of the intrinsic Quality Control
coagulation mechanism(5). Rel iabil ity of test results should be monitored within each run using
Coagulation Control Plasmas. Each laboratory should establish a control range
Kit Components to determine the al lowable variation in day to day performance of each control
Reagent/ Product Code Description plasma. Agappe Coagulation Control (Bi-Level) (Product Code: 11624003) is
Component 12602003 available.
APTT Reagent 5 x 4 mL Rabbit brain cephalin, ellagic acid Calculation of Results
For best results, duplicate samples are recommended. APTT results should be
Calcium chloride 5 x 4 mL Calcium chloride (0.02 M)
reported as clotting time in seconds. Calculate the mean clotting time of
Accessory Kit 200 nos Coagulation Cuvette with Steel Beads duplicate samples and controls. Differences between duplicate results should
(provided in separate
1 pc Forceps be less than 5%. Repeat the test if necessary.
box)

Risk & Safety Limitations

To prevent discrepant results ensure the blood to anticoagulant ratio is 9:1.
Material Safety data sheets (MSDS) will be provided on request
Grossly lipemic or hemolysed samples may produce erroneous APTT values(6).
Reagent Delay in testing, difficulty in specimen col lection, or venipucture above the
The APTT reagent is a preparation of rabbit brain cephalin and el lagic acid site of a heparin lock may result in falsely prolonged APTT results(7). The APTT
activator with buffer, stabilizers and preservatives. The reagent is provided ready may also be influenced by certain drugs and medications(8). APTT results can
to use. vary with anticoagulation therapy depending upon the type and dosage of
Precautions anticoagulant, the route of administration and the time of administration of
Do not ingest. Avoid contact with skin, eyes or clothing. the last dose.

Waste Management Expected Values

Reagents must be disposed off in accordance with local regulations. APTT results are influenced by the method of clot detection and can vary from
laboratory to laboratory. In general an APTT test performed on a photo-optical
Storage and Stability
coagulometer wil l give clotting time for normal plasma in the range of 24 to 39
The APTT reagent is stable to the expiry date shown on the label when stored in seconds. Therapeutic ranges for monitoring oral anticoagulation therapy wil l
the original container at 2 to 8OC. vary from laboratory to laboratory, therefore it is essential that each laboratory
Specimen Col lection and Preparation establish relevant APTT ranges for its respective patient population.
Test plasma should be prepared from citrated whole blood without heparin, EDTA Abnormal results obtained with a plasma from a patient not on anticoagulant
or oxalate. therapy may indicate a factor deficiency or the presence on an inhibitor. The
1. Blood Col lection using Syringe Method: Draw venous blood into a plastic result may also be due to the effects of certain drugs and medications. Additional
or siliconized syringe. Immediately transfer 9.0 mL of blood into a tube procedures such as the PT test and mixing studies using factor deficient plasma
containing 1.0 mL of 3.2% or 3.8% sodium citrate solution. are usual ly required.
2. Blood Col lection using an Evacuated Blood Col lection Tube: Draw venous Performance Characteristics
blood into a commercial vacuum tube containing 3.2% or 3.8% sodium citrate Precision:
solution. Insure that a full draw has been obtained since the ratio of 9 parts Within-run precision was assessed using normal and abnormal plasma controls.
blood to 1 part citrate is critical. A heparinized lock or transfer line should not
be used. It is generally recommended that the second or third tube draw be
used for coagulation tests. Sample Mean SD CV%
3. Plasma Preparation: Mix well by inversion and centrifuge at 2,500 x g for 15 Control 1 22.5 1.32 5.85
minutes soon after blood col lection. Unless samples are to be processed
Control 2 55.62 2.11 6.34
immediately, transfer the plasma into a plastic tube. Plasma that is clearly
hemolyzed or contains > 10,000 platelets per cubic milliliter or red cells is not
suitable for coagulation testing. Comparison
4. Plasma Storage: Plasma samples should be transferred to a plastic tube as A comparison study has been performed between Agappe reagent and another
soon as possible and stored refrigerated (2 to 8oC). Plasma samples should be internationally available reagent yielded a correlation coefficient of r2= 0.8902 and
tested within 4 hours and should not be incubated at 37OC for more than 5 a regression equation of y = 0.9383x + 0.9387.
minutes to avoid loss of factors V and VII.

‘Agappe Hills’, Dist. Ernakulam, Kerala, India-683 562.

Tel. +91 484 2867 000 | Customer Support No.: 1800 425 7151(Toll Free) ISO 9001:2015
[email protected] | www.agappe.com REV. NO.: ADL/IFU/APTT/CLG/R03 EN ISO 13485:2016
 200 Tests
APTT ‘New’ 12602003

References 4. Hougie C, The Biochemistry of Blood Coagulation, In Laboratory Evaluation of

1. Human Blood Coagulation, Hemostasis and Thrombosis, 3rd ed. R Biggs, CR Coagulation, American Society of Clinical Pathologists Press, Chicago p 2 (1982)
Rizza, Editors, Blackwell Scientific Publications, London (1984) 5. Owen CA, Bowie EJW, Thomson JH, The Diagnosis of Bleeding Disorders, Little
2. Cole E, Hall ER, Wu KK, Principles of Antithromboltic Therapy. In Wu KK, Brown and Company, Boston p 110 (1975)
Thomboembolic Disorders, PSG Publishing Co. Inc., Litteton, p 91 (1984) 6. Harker LA, Hemostasis Manual, FA Davis Co, Philadelphia p 62(1974)
3. Triplett DA, Heparin: Clinical use and Laboratory Monitoring. In Triplett DA, 7. Triplett DA, Harms CS, Procedures for the Coagulation Laboratory, American
Laboratory Evaluation of Coagulation, American Society of Cl inical Society of Clinical Pathologists Press, Chicago, p 7 (1981)
Pathologists Press, Chicago p 272 (1982) 8. Young DS, Pestaner LC, Gibberman V, Effects of Drugs on Clinical Laboratory
Tests, Clin Chem 21; 1D (1975)

‘Agappe Hills’, Dist. Ernakulam, Kerala, India-683 562.

Tel. +91 484 2867 000 | Customer Support No.: 1800 425 7151(Toll Free) ISO 9001:2015
[email protected] | www.agappe.com REV. NO.: ADL/IFU/APTT/CLG/R03 EN ISO 13485:2016