BIOLABO

www.biolabo.fr
BIO-TP Prothrombin Time (PT)
For determination of Prothrombin Time (INR) in human plasmas
MANUFACTURER:
BIOLABO SAS,
REF 13885 R1 10 x 2 mL R2 1 x 25 mL
Les Hautes Rives
02160, Maizy, France REF 13880 R1 6 x 4 mL R2 1 x 25 mL
REF 13881 R1 6 x 12 mL R2 1 x 80 mL

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TECHNICAL SUPPORT AND ORDERS
Tel : (33) 03 23 25 15 50
IN VITRO DIAGNOSTIC USE
Fax: (33) 03 23 256 256

CLINICAL SIGNIFICANCE (1) (6) (7) REAGENTS PREPARATION

The Prothrombin time (PT) is a useful basic coagulation screening test
to investigate the extrinsic coagulation pathway. · Thromboplastin (Vial R1)
PT (in sec.) converted into PT (%) allows the evaluation of the Use a non-sharp instrument to remove aluminium cap from the vial.
prothrombinic activity, referring to a normal plasma (100 %). ● Reconstitution buffer (Vial R2): Ready to use
A deficient prothrombinic activity is found in the following clinical ● Working reagent: Once opened, add promptly to the contents of
states: the vial R1 the amount of dilution buffer (vial R2) stated on the label.
· Hemorrhagic disease of the newborn. Mix gently until complete dissolution
· Liver failure (cirrhosis, hepatitis...).
· Vitamin K deficiency or treatment with vitamin K antagonists. STABILITY AND STORAGE
· Congenital deficits in one of the factors associated with the Stored away from light, well caped in the original vial at 2-8°C,
prothrombinic complex, real prothrombin (factor II), proaccelerin and used as described, reagents are stable:
(factor V), proconvertin (factor VII) and Stuart’s factor (factor X)
· Circulating anticoagulants Unopened:
· Fibrinolysis · Until expiry date stated on the label of the kit.
· DIC (disseminated intravascular coagulation). Once reconstituted:
Monitoring of treatment with vitamin K antagonists: · Transfer requested quantity, well recap vials and store at 2-8°C
The PT (in sec.) may be converted into INR (International Normalised · Working Reagent (R1+R2) is stable for:
Ratio). In that case, the origin of the thromboplastin has no incidence ü 8 hours at room temperature
on the determination of the expected values. An international ü 5 days at 2-8°C.
standardisation about INR reference intervals has been established for · Don’t use working reagent after expiry date.
treatment and prophylaxis of venous and arterial thromboembolisms.
Avoid results in INR in the case of pre-operative check-up or SPECIMEN COLLECTION AND HANDLING (2) (8)
investigations for liver diseases. Careful venipuncture.
· Blood/anticoagulant ratio: 4.5 mL of blood for 0.5 mL of sodium citrate
PRINCIPLE (4) 2 H2O 0.109 M. Avoid blood drawing with a syringe that could result
Quick and al. method. Principle as follows: in the formation of micro-clots. Centrifuge for 10 minutes at 2500 g.
The clotting time is measured at 37°C in the presence of tissular · Run the assay within 4 hours after collection, storing plasma at room
thromboplastin and calcium. The PT (in sec.) so measured is temperature (15-25°C).
converted into PT (%) or INR. · Collection on citrate Hepes tube increases the specimen stability up
to 8 hours.
REAGENTS
R1 Thromboplastin REAG LIMITS (2) (3)
Freeze-dried Thromboplastin (Rabbit cerebral tissue) Contamination by Thromboplastin or hemolysed specimens may also
shorten the result of PT (in sec.).
R2 Reconstitution Buffer BUF For a more comprehensive review of factors affecting this assay, refer
HEPES Buffer, Stabilizer to the publication of Young D.S.
According to 1272/2008 regulation, these reagents are not classified
MATERIAL REQUIRED BUT NOT PROVIDED
as dangerous
1. Basic medical analysis laboratory equipment.
2. Automated or semi-automated coagulation analyzer
SAFETY CAUTIONS
3. REF 13883 Owren Koller buffer to establish the Thivolle line (in %).
BIOLABO reagents are designated for professional, in vitro diagnostic 4. Graph paper
use (do not pipette with mouth).
· Refer to current Material Safety Data Sheet available on request or
on www.biolabo.fr
· Verify the integrity of the contents before use.
· Waste disposal: Respect legislation in force in the country.
All specimens should be handled as potentially infectious, in
accordance with good laboratory practices using appropriate
precautions. Respect legislation in force in the country.

IVD REF LOT ®

Manufacturer Use by In vitro diagnostic Temperature limitation Catalogue number See insert Batch number Store away from light sufficient for dilute with

H2O
Demineralized water Biological hazard

Made in France Latest revision: www.biolabo.fr Revision: 23/01/2018

CALIBRATION EXPECTED VALUES (2) (6) (9)

INR with MNPT and ISI (all methods) Normal PT (in sec.):
ISI value of the reagent is determined by testing human plasmas with this Usually between 11 and 16 seconds (depending on the reagent).
Thromboplastin and with an Internal Reference Thromboplastin traceable Normal PT (%): Ranging between 70 to 100 %.
to RBT16 (WHO International Standard Thromboplastin, Rabbit plain). Values over 100 % are considered as normal.
Obtained PT (sec values) with the 2 thromboplastins are plotted on log to Oral anticoagulant therapy (OAT):
log graph and the slope is calculated. Therapeutic range in INR PT (%)
The ISI of the Reagent is calculated multiplying the slope by ISI of the Indications Rabbit
Target Acceptable range thromboplastin
Internal Reference Thromboplastin.
Pre-operative and during surgery:
Reference plasma (MNPT): Prepare a pool of freshly collected normal *Hip surgery 2.5 2.0 – 3.0 35 %
plasmas. Measure in triplicate the clotting time and calculate the mean.
*Other surgery 2.0 1.5 – 2.5 40 %
Determine patients results in INR using MNPT (Mean normal plasma Venous thrombosis prophylaxis 2.5 2.0 – 3.0 35 %
time) and ISI of the reagent (International Sensitivity Index) indicated in Evolutive phlebitis, pulmonary
the specific batch table (§CALCULATION). embolism, recurrent phlebitis
3.0 2.0 – 4.0 27 %

INR and PT% with TP-CALSET REF 13965 Arterial prophylaxis, mechanical 3.5 3.0 – 4.5 25 %
prosthetic valves
· Automated System SOLEA 100: Perform a calibration with TP-
CALSET (3 different levels)
PROCEDURE
· Manual procedure on Semi-automates BIO SOLEA 2, BIO SOLEA 4
Automated procedure on SOLEA 100:
Determine in triplicate the clotting time of each level of TP-CALSET
Full detailed specific application available on request
Manual procedure on semi-automates BIO SOLEA 2, BIO SOLEA 4:
QUALITY CONTROL Pre-incubate Thromboplastin at least 15 minutes at 37°C and mix gently.
REF 13961 Control Plasma level 1 6 x 1 mL Plasma 0.1 mL
REF 13962 Control Plasma level 2 6 x 1 mL Incubate for 2 minutes at 37°C
Working reagent (R1+R2) homogenised at 37°C 0.2 mL
REF 13963 Control Plasma level 3 6 x 1 mL
Or The automatic countdown timer will start immediately after working
reagent addition and stop when the clot is formed.
REF 13971 Coatrol 1 6 x 1 mL
Note:
REF 13972 Coatrol 2 6 x 1 mL · Performances and stability data have been validated on SOLEA 100
and Thrombolyzer Compact X (available on request).
· External quality control program.
· With manual procedure and on other automated coagulation analyzer,
It is recommended to control in the following cases: performances and stability data must be validated by user.
· At least once a run. · Other validated applications or proposal are available on request.
· At least once within 24 hours.
· When changing vial of reagent.
· After maintenance operation on the instrument. CALCULATION (6)
If control is out of range, apply following actions: INR with MNPT and ISI (all methods):
1. Repeat the test with the same control plasma. ISI
2. If control is still out of range, prepare fresh control plasma and repeat
the test.
INR calculation = ( Patient’s time
Mean normal time
)
3. If control is still out of range, calibrate with a new vial of reagent. · SOLEA 100 automate and BIO SOLEA2, BIO SOLEA4 semi
4. If control is still out of range, use a new vial of reference plasma and automate: INR results are calculated automatically after input in the
repeat the test. system
5. If control is still out of range, please contact BIOLABO technical support
· Manual Procedure: Refer to the enclosed calculation board (including
or your local Agent.
ISI value) corresponding to the current batch number to calculate PT
(%) and INR. Select the column corresponding to the measured time for
PERFORMANCE AT 37°C ON SOLEA 100 MNPT. Identify the patient’s PT (sec) in this column. On the same line,
Studies realised on normal and pathological plasmas
refer to the corresponding PT (%) or INR.
Within Between
run Level 1 Level 2 Level 3 run Level 1 Level 2 Level 3 Or plot on a paper graph the Thivolle line (using dilution of normal
N = 20 N = 20 plasma pool in Owren Koller buffer REF 13883). Plot the clotting time
Mean 89 36 22 Mean 89 35 21 measured for the patient on the Thivolle line and then read on the X-
(%) (%)
axis the reciprocal dilution corresponding. Reverse and multiply by 100
S.D. (%) : 1,3 0,57 0,40 S.D. (%) : 1,66 1,47 0,64
to obtain the PT (%) for patient.
C.V. %: 1,46 1,57 1,85 C.V. %: 1,86 4,17 3,08 INR and PT% with TP-CALSET REF 13965
Comparison with commercially available reagent (same method): · SOLEA 100 automate: INR and % results will be calculated
automatically according to calibration curve
170 plasmas located between 14% and 110% were tested with the 2
reagents on BIO SOLEA4 coagulometer: · Manual Procedure on semi automate BIO SOLEA2 et 4:
Enter the mean of the clotting time found for each level of TP-CALSET
y = 1,0287 x +0,1601 r= 0,9863
and the corresponding PT % in the system. PT % results will be
Interferences on PT (sec, INR): calculated automatically according to calibration curve
Turbidity No interference up to 0,404 abs
Low Molecular REFERENCES
Positive interference from 0.11 IU anti Xa
weight heparin
(1) Caen J., Larrieu MJ, Samama M : « L’hémostase. Méthodes d’exploration et
Bilirubin Positive interference from 171 µmol/L diagnostic pratique » Paris : L’Expension Scientfique, p.344-347, (1975).
th
Hemoglobin No interference up to 258 µmol/L (2) Clinical Guide to Laboratory Test, 4 Ed., N.W. TIETZ (2006) p.928-929
Other substances may interfere with the results (see § Limits) (3) YOUNG D.S., Effect of Drugs on Clinical laboratory Tests, 4th Ed. (1995)
Onboard stability: at least 24 hours when kept 8 hours per days onboard p.3-513 à 3-517
(4) Quick A. J.- J. Am. Assoc., (1938), 110,p. 1658-1662
Calibration Stability: 6 weeks (5) Duckert F., Marbet G.A. - Méd., et Hyg., (197)7, 35, p. 911
Make a new calibration when changing reagent batch, if quality control (6) Goguel A.F. - Feuillets de Biologie, (1985), 36, (146) p. 25-28.
results are found out of the established range and after maintenance (7) Houbouyan-Reveillard et al. Spectra biologie (2003) vol.22, n°132 p.33-37
(8) Neofotistos D, Oropeza M., Ts’ao C-H : « Stability of plasma for add-on PT
operations. and PTT tests » Am. J. Clin. Pathol. 109, 6, 758-763, (1998).
(9) Sampol J., Arnoux D., Boutière B. : « Manuel d’hémostase » Paris, Ed. Elsevier,
147-163 (1995)

Made in France Latest revision: www.biolabo.fr Revision: 23/01/2018