PT-HS

CONTENTS
PT-HS
Prothrombin Time (PT)
REF 3510101 PT-HS 10 x 5 mL
PT with Calcium
High-Sensitivity Reagent
For in vitro diagnostic use only

PRINCIPLE 3. Plasma Storage

o
Plasma samples may be stored at 18 to 26 C for up to 2 hours; at 2
The capacity of blood to form a fibrin clot by means of the to 8
o
C for up to 4 hours; frozen at –20
o
C for up to 2 months or at –
extrinsic hemostatic pathway requires thromboplastin, calcium, o
70 C for up to 6 months. Plasma may be re-centrifuged prior to
(4,5)
factors I, II, V, VII and X . The PT-HS reagent provides a freezing to assure that all cells are removed. Quick thaw frozen
source of tissue thromboplastin and calcium that specifically samples and test immediately. The samples must not have any
activates factor VII in the extrinsic coagulation pathway. The o
contact with glass. Do not incubate samples at 37 C for longer than
factors involved in the intrinsic coagulation pathway are 5 minutes to avoid the loss of factors V and VII. The loosing of
6
bypassed . factor V can prolong the PT.
Therefore, deficiencies of intrinsic pathway factors (VIII, IX and
XII) are not detected using the PT test.
INTERFERENCES
REAGENT COMPOSITION
− PT clotting times may be prolonged by substances including
PT-HS Lyophilized thromboplastin of rabbit brain and CaCl2. corticosteroids, EDTA, oral contraceptives, asparaginase,
Sodium azide 0.05%. clofibrate, erythromycin, ethanol, tetracycline and
International Sensitivity Index (ISI): 1.07 – 1.39. anticoagulants such as heparin and Coumadin.
− PT may be shortened by substances including antihistamines,
DIL PT Diluent butabarbital, caffeine, oral contraceptives, phenobarbital and
10
Optative. Plasma Control Level 1 Ref. 3520101 vitamin K .
Plasma Control Level 2 Ref. 3520201.
ADDITIONAL EQUIPMENT
STORAGE AND STABILITY
− Coagulometer or stopwatch and bath at 37ºC ± 0.5ºC.
Store at 2-8ºC. The reagents are stable until the expiry date
− General laboratory equipment.
stated on the label. The reconstituted PT-HS reagent is stable for 8
o
hours at 37ºC and 5 days stored in the original container at 2-8 C.
Do not freeze. PROCEDURE

REAGENT PREPARATION This procedure pertains to manual or semi-automated coagulation

systems. Refer to your instrument manual for more detailed
1. Reconstitute the contents of PT-HS with one vial of PT Diluent. instrument specific instructions.
2. Replace the stopper and thoroughly mix the vial contents. Let
o
stand for no less than 30 minutes prior to use to assure 1. Pre-incubate the reconstituted PT reagent to 37 C for at least
complete hydration of the contents. 10 minutes.
2. Pipette 50 μL of sample or control plasma into a test cuvette
SAMPLES 3. Incubate at 37oC for 2 minute.
4. Rapidly add 100 μL of the pre-incubated PT reagent,
Test plasma should be prepared from citrated whole blood without simultaneously starting the timer.
heparin, EDTA or oxalate. 5. 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 9.0 mL of blood into a tube containing 1.0
mL of 3.2% or 3.8% sodium citrate solution. or, CALCULATIONS
- Draw venous blood into a commercial vacuum tube containing
3.2% or 3.8% sodium citrate solution. Calculate the mean clotting time of duplicate samples and controls.
Insure that a full draw has been obtained since the ratio of 9 parts Differences between duplicate results should be less than 5%.
blood to 1 part citrate is critical. A heparinized lock or transfer line Repeat the test if necessary.
should not be used. It is generally recommended that the second or The PT result may be reported as 4 different ways:
third tube draw be used for coagulation tests.
1. Seconds, observed clotting time.
2. Plasma Preparation
- Mix well by inversion and centrifuge at 2,500 x g for 15 minutes 2. PT ratio (PR):
soon after blood collection. Unless samples are to be processed
immediately, transfer the plasma into a plastic tube. Plasma that PT of the patient in sec
is clearly hemolyzed or contains > 10,000 platelets per cubic PT ratio (PR) =
PT of normal plasma in sec
millilitre or red cells is not suitable for coagulation testing.

QUALITY SYSTEM CERTIFIED LINEAR CHEMICALS S.L. Joaquim Costa 18 2ª planta. 08390 Montgat, Barcelona, SPAIN
ISO 9001 ISO 13485 Telf. (+34) 934 694 990 Fax. (+34) 934 693 435. website www.linear.es
 CLINICAL SIGNIFICANCE
3. Percent activity.
The PT reagent is an in vitro diagnostic assay intended for use in
4. International Normalized Ratio (INR) performing the one stage prothrombin time (PT) test and assays
The INR is recommended for use with patients undergoing anti- which are based on a modified prothrombin time.
coagulation therapy. The prothrombin time is the method of choice for monitoring oral
The International Committee for Standardization in Hematology and 1
anticoagulation therapy and is a fundamental screening test for
the International Committee on Thrombosis and Hemostasis have acquired or inherited bleeding disorders. During oral anti-
agreed on recommendations for the reporting of Prothrombin Time coagulation therapy, the activity of vitamin K-dependant clotting
results as an International Normalized Ratio (INR). The INR is factors (II, VII, IX, X, Protein C and Protein S) is reduced and PT
based on the International Sensitivity Index (ISI) of Thromboplastin time is increased. The test is used for quantitative determination of
7-9
reagents . blood clotting factors in the extrinsic (VII) and common pathways
PT reagents are assigned with an ISI value by calibration against 2,3
(II, V and X) of coagulation .
an International Reference Preparation (IRP 67/40) with an
assigned ISI of 1.0. The ISI value for each lot of PT reagent
ANALYTICAL PERFORMANCE
appears on the vial label.
The INR is calculated using the following formula: - Linearity: INR 0.8 – 5.0.
ISI - Sensitivity range: ISI : 1.07 – 1.39.
INR = (Patient PT / Mean Normal PT)

ISI = Specific International Sensitivity Index for the - Normal PT time mean: 12.0 – 14.7 sec.
Reagent/Instrument system. The ISI is lot specific.
- Analytical sensitivity: 3%.
Mean Normal PT = Lot specific mean of the normal range, as
determined by each laboratory for the Reagent /Instrument System. - Traceability: 2 international reference laboratories of WHO , by
It is usually based upon the PT mean plus or minus 2 to 3 standard Standard Rabbit Plain Thromboplastin (WHO Thrombosis Center
deviations using 20 or more individuals. Leiden, and Univ. Ontario, Canada).
REFERENCE VALUES - Accuracy: Results obtained with this reagent did not show
PT results are influenced by the method of clot detection and can significative differences when compared with reference reagents.
vary from laboratory to laboratory. In general, PT tests performed Details of the comparison experiments are available on request.
with normal plasmas will give clotting times in the range of 13 to 15
seconds on a photo-optical coagulometer, from 11 to 15 seconds if REFERENCES
a mechanical coagulometer is used, and between 12 and 15 1. Deykin, D, Anticoagulant therapy. In: Colman, R.W., Hirsh, J, Marder,
seconds if the manual method is applied. However, each laboratory V., Salzman, EW (Eds.); Heamostatis and Thrombosis, JB Lippincott,
should establish a normal range using individuals representative of Philadelphia, p1000 (1982).
its patient population. A new normal range should be established 2. Errichette AM, Holden A, Ansell J; Management of Oral Anticoagulant
with any change of instrumentation, blood collection techniques, or Therapy: experience with an Anticoagulation Clinic, Arch. Inter.
anticoagulant. A Mean Normal PT range should be reestablished or Medicine 144; p1966 (1984).
verified when changing lots of the same reagent .
(11) 3. Hirsh J, Dalen JE, Deykin D., Polter L; Oral Anticoagulants:
Mechanisms of Action, Clinical Effectiveness and Optimal Therapeutic
Therapeutic ranges for monitoring oral anticoagulation therapy will Range, Chest 102 (suppl):312S, (1992).
vary from laboratory to laboratory. Therefore, it is essential that 4. Miale JB; Laboratory Medicine-Hematology, 4th edition, CV Mosbe, St
each laboratory establish relevant PT ranges for its respective Louis, (1972).
patient population. 5. Furie B, Furie BC; Molecular and Cellular Biology of Blood
Abnormal results obtained with a plasma from a patient not on Coagulation, N Eng J Medicine 326;p800 (1992).
anticoagulant therapy may indicate a factor deficiency or the 6. Hougie C; The Biochemistry of Blood Coagulation; In Triplett DA,
presence of an inhibitor. The result may also be due to the effects Laboratory Evaluation of Coagulation, American Society of Clinical
(12) Pathologists Press, Chicago, p2 (1982).
of certain drugs and medications . Additional procedures such as 7. WHO Expert Committee on Biological Standardization, 33 Report.
an APTT and mixing studies using factor deficient plasma are Technical Report Series 687, WHO, Geneva (1983).
usually required. 8. Kirkwood T; Calibration of Reference Thromboplastins and
Standardization of the Prothrombin Time Ratio, Thromb Haemostasis
QUALITY CONTROL 49; p238 (1983).
9. International Committee for Standardization in Haematology and
Control sera are recommended to monitor the performance of International Committee on Thrombosis and Haemostasis. Amer J Clin
assay procedures. They should be used as sample. Path 88; p779 (1985).
10. Young DS, Thomas DW, Friedman RB, et al.; Effect of Drugs in
REF 3520101 PLASMA CONTROL LEVEL 1 Clinical Tests, Clin Chem 18; p1041 (1972).
11. National Committee for Clinical Laboratory Standards: One-Stage
REF 3520201 PLASMA CONTROL LEVEL 2 Prothrombin Time (PT) Test and Activated Partial Thromboplastin Time
(APTT) Test, NCCLS Document H47-A (1996).
Each laboratory should establish its own Quality Control scheme
12. Laposala M, Connor A, Hicks D, Phillips D: The Clinical Hemostasis
and corrective actions if controls do not meet the acceptable handbook, Year Book Medical Publishers Inc. (1989).
tolerances.

C35101-2/0802
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QUALITY SYSTEM CERTIFIED LINEAR CHEMICALS S.L. Joaquim Costa 18 2ª planta. 08390 Montgat, Barcelona, SPAIN
ISO 9001 ISO 13485 Telf. (+34) 934 694 990 Fax. (+34) 934 693 435. website www.linear.es
 PT

MASTER CURVE

Introduzca los siguientes valores en el coagulómetro

Enter the following values in the coagulometer

LOT Nº 35025

Expiry date 2010-08

g/L SEC.

100% 13,70 SEC

50% 21,93 SEC

33% 30,41 SEC

25% 38,39 SEC

ISI 1.07

*Print, and archive the calibration line. This calibration curve is valid for this lot of PT only.

C35103-2/0802
R1.cas

QUALITY SYSTEM CERTIFIED LINEAR CHEMICALS S.L. Joaquim Costa 18 2ª planta. 08390 Montgat, Barcelona, SPAIN
ISO 9001 ISO 13485 Telf. (+34) 934 694 990 Fax. (+34) 934 693 435. website www.linear.es