In Vitro

Diagnostic

BE PT LI Thromboplastin low ISI REF 771100: RE (5 x 5 mL), DIL (2 x 15 mL)

Reagent for determination of Prothrombin time (PT) in human plasma REF 771101: RE (8 x 12 mL), DIL (8 x 12 mL)
PRINCIPLE (4) PROCEDURE
Quick method: tissue thromboplastin and calcium are added to citrated plasma, Manual method on semi-automated systems
activating the factors of extrinsic coagulation pathway. The clotting time (time to Pre-incubate reagent 15 min to reach a temperature of 37 °C and mix gently before use:
formation of a fibrin clot) is measured at 37 °C. • Plasma: 100 µL
C L I N I C AL S I G N I F I C AN C E ( 1 ) ( 6 ) ( 7 ) Incubate for 120 sec at 37 °C
• Thromboplastin (37 °C): 200 µL
The Prothrombin Time is used as a screening test to detect disorders in the extrinsic
The automatic countdown timer will start immediately after Thromboplastin addition and
coagulation pathway. stop when the clot is formed.
The results in seconds are converted into percentages to evaluate the prothrombinic Automated method on Behnk Thrombolyzer series
activity in comparison to a reference plasma consider as 100%. Refer to the full detailed application specific to the automated system.
A deficient prothrombinic activity can occur in the following cases: Note:
Hepatic disease; Hemorrhagic disease of new born; Congenital factor deficiency (factors • Performances and stability data have been validated on Thrombolyzer Compact X
II, V, VII or X); Vitamin K deficiency; Vitamin K antagonist treatment; Circulating (available on request).
anticoagulants; Fibrinolysis; Disseminated intravascular coagulation (DIC) • With manual procedure and on other automated coagulation analyzer, performances and
The results in seconds are converted into INR (International Normalized Ratio) in the stability data must be validated by user.
particular use of monitoring oral anticoagulant therapy (Vitamin K antagonists). • Other validated applications or proposal applications are available on request.
R E AG E N T S C AL I B R A T I O N
RE PT LI Reagent PT INR and % with Calibrator Set
Thromboplastin (lyophilized from rabbit brain). Use calibrator set REF 775200: BE Cal Set traceable to RBT16 (WHO International
According to 1272/2008 regulation, this reagent is not classified as dangerous.
Standard Thromboplastin, Rabbit plain).
• Automated method on Behnk Thrombolyzer series:
DIL PT Diluent Diluent Attention
Perform a Calibrator Set calibration with BE Cal Set.
Hepes buffer, calcium.
• Manual method on semi-automated systems (PT %): Prepare a calibration curve with
Skin Sens.1: H317 - May cause an allergic skin reaction Cal1, Cal2 and Cal3. Measure in triplicate the clotting time of each level.
P261: avoid breathing sprays, P280: Wear protective gloves/protective clothing/eye
protection/face protection, P302+352: IF ON SKIN: Wash with soap and water. PT INR with MNPT and ISI (all methods)
P333+313: If skin irritation or a rash occurs: Get medical advice/attention, • MNPT (Mean Normal Prothrombin Time)
P501: Dispose of contents/container in accordance with dangerous goods regulations To determine the MNPT prepare a pool of freshly collected normal plasmas. Measure in
Classification due to: Nickel Sulfate < 1%. For more details refer to current Material Safety Data triplicate the clotting time and calculate the mean.
Sheet (MSDS) • ISI (International Sensitivity Index): Refer to the batch specific table.
Once reconstituted: Working Reagent (vial RE) is classified as Buffer (vial DIL) (6)
C AL C U L AT I O N
S AF E T Y C AU T I O N S
PT INR and PT % with Calibrator Set
Behnk reagent kits are designated for professional in vitro diagnostic use.
• Automated method on Behnk Thrombolyzer series:
Good Laboratory Practices must be applied during use of reagents, reference or control
PT INR and PT % will be calculated automatically according to two calibration curves.
plasmas, and human samples and should be handled as potentially infectious.
• Semi-automated systems: Enter the mean of the clotting time found for each BE Cal
For further information, Material Safety datasheet is available upon request.
Set plasma and the corresponding PT % in the system. PT % concentration will be
Dispose of waste in accordance with the local regulations.
calculated automatically according to calibration curve.
P R E P AR AT I O N O F R E AG E N TS
PT INR with MNPT and ISI (all methods)
RE: Reconstitute the lyophilisate with the amount of DIL indicated on the label of RE. The MNPT and ISI are to be used to calculate result in INR.
Cap vials and mix gently the RE vial until complete dissolution. • Calculate INR as follows: INR = (Patient time / MNPT) ISI
DIL: Ready for use • Refer to enclosed batch specific table, selecting the suitable column according the
S T AB I L I T Y AN D S T O R AG E MNPT. Identify the raw corresponding to patient’s result and refer to the result in PT
Unopened vials stored at 2-8 °C are stable until the expiry date stated on the label. % and PT INR indicated in column “%” and “INR”.
RE: after reconstitution the working reagent is stable 5 days at 2-8 °C, or 6 h at 37 °C. • For semi-automated systems and Behnk Thrombolyzer series the INR will be
DIL: once opened, if stored at 2-8 °C and free from contamination, contents of the vial is calculated automatically after input in the system.
stable until the expiry date stated on the label. P E R F O R M AN C E S
Do not use any reagent after expiry date. The within run and between run studies were performed with normal and abnormal
S AM P L E S C O L L E C T I O N AN D H A N D L I N G ( 2 ) ( 8 ) plasma on Thrombolyzer Compact X.
Plasma from careful venipuncture with anticoagulant ratio of 1/10 (trisodium citrate Within run Normal High Between run Normal High
solution 0.109 M). Mix immediately the blood with anticoagulant. N = 20 Plasma Plasma N = 20 Plasma Plasma
Avoid drawing with a syringe that could result in the formation of micro-clots. Mean (%) 96.6 30.3 Mean (%) 96 30.4
Centrifuge 10 minutes at 2500 g. S.D. (%) 0.98 0.54 S.D. (%) 1.81 0.99
The specimen is stable 4 hours after collection, at room temperature (15-25 °C). C.V. % 1.01 1.77 C.V. % 1.88 3.26
Collection on Citrate Hepes tube increases the sample stability to 8 hours. Comparison with commercially available reagent, same method: 167 plasmas located
LIMITS (2) (3) between 14% and 110%:
y = 1.1376x – 1.4301 r= 0.9958
Samples contaminated by thromboplastin or hemolysis may shorten the clotting time.
Interferences (sec, INR):
For a more comprehensive review of factors affecting this assay, refer to the publication
of Young D.S. Turbidity No interference up to 731 mg/dL of Triglycerides
Low Molecular weight heparin Positive interference from 0.114 IU anti Xa
M AT E R I AL R E Q U I R E D B U T N O T P R O V I D E D Unfractionated heparin Positive interference from 0.038 IU anti Xa
Basic medical analysis laboratory equipment Bilirubin Positive interference from 228 µmol/L
Automated or semi-automated Coagulation analyzer Hemoglobin No interference up to 240 µmol/L
REF 050813: Magnetic stirrers 8 x 1.5 mm, for Behnk Thrombolyzer series. Other substances may interfere with the results (see § Limits)
E X P E C T E D V AL U E S ( 2 ) ( 6 ) Calibration Stability: Make a new calibration when changing reagent batch, if quality
PT (sec): Normal values between 11 and 16 sec. control results are found out of the established range and after maintenance operations.
PT (%): Normal values 70 % to 100 %. Values over 100 % have no clinical significance. REFERENCES
PT (INR): It is advised to each laboratory to establish its own reference range of (1) Caen J., Larrieu MJ, Samama M : « L’hémostase. Méthodes d’exploration et diagnostic pratique »
expected values. Paris : L’Expension Scientfique, p.344-347, (1975).
(2) Clinical Guide to Laboratory Test, 3 rd Ed., N.W. TIETZ (1995) p.526-529
Q U AL I T Y C O N T R O L
(3) YOUNG D.S., Effect of Drugs on Clinical laboratory Tests, 4 th Ed. (1995) p.3-513 à 3-517
REF 773100: BE Trol 1; REF 773101: BE Trol 2 (4) Quick A. J.- J. Am. Assoc., (1938), 110,p. 1658-1662
Controls are required for checking the accuracy and reproducibility of the results. (5) Duckert F., Marbet G.A. - Méd., et Hyg., (1977), 35, p. 911
(6) Goguel A.F. - Feuillets de Biologie, (1985), 36, (146) p. 25-28.
The control intervals should be adapted to each laboratory’s individual requirements.
(7) Houbouyan-Reveillard et al. Spectra biologie (2003) vol.22, n°132 p.33-37
Values obtained should fall within the defined limits. Follow the applicable government (8) Neofotistos D, Oropeza M., Ts’ao C-H : « Stability of plasma for add-on PT and PTT tests » Am. J.
regulations and local guidelines for quality control. Clin. Pathol. 109, 6, 758-763, (1998).

IVD REF LOT  H2O

Manufacturer Use by In Vitro Diagnostic Temperature limitation Catalogue number See insert Batch number Store away from light Sufficient for Dilute with Demineralized water Biological hazard

Manufactured by Distributed by
BIOLABO S.A.S. Kommanditgesellschaft Behnk Elektronik GmbH & Co. T. +49 (0)40-529 861 0
Les Hautes Rives Hans-Böckler-Ring 27 F. +49 (0)40-529 861 99
02160 Maizy, France 22851 Norderstedt, Germany info@behnk.de
www.behnk.de
Made in France Latest revision : www.behnk.de Revision : 20171219