TM

2 x 4 mL
APTT 12602001

Intended Use Procedure

This reagent is intended for the in vitro determination of Activated Partial APTT of each sample should be determined at each level twice. This procedure
Thromboplastin Time (APTT) in citrated plasma. pertains to manual or semi-automated coagulation systems. Refer to your
instrument manual for more detailed instrument specific instructions.
Cl inical Significance
Bring contents of the vial to room temperature and then swirl gently to mix to a
The activated partial thromboplastin time (APTT) is used as a general screening
homogenized suspension. Keep the reagents at 37OC for 10 minutes prior to use.
test for the detection of coagulation abnormalities in the intrinsic pathway. APTT
is sensitive to deficiencies or abnormalities of factors VIII, IX, XI, XII, X, V, II and 1. Gently swirl the reagent vials before use. Do not shake.
I. APTT is also sensitive to inhibitors of blood coagulation such as lupus inhibitor 2. Pre-warm enough volume of Reagent 1 (CaCl2) for immediate use, in a clean &
and fibrin/fibrinogen degradation products. APTT is the most widely used method dry plastic test tube at 37 OC.
for monitoring intravenous heparin anticoagulation therapy.
3. Pipette 100 µL of test plasma or control in to a test cuvette at 37 OC.
Principle
4. Pipette 100 µL of the pre-warmed reagent 2 (APTT Reagent) in to the test
In presence of Calcium ions cephaloplastin activates coagulation factors of intrinsic
cuvette.
pathway in plasma lead ing to clot formation. Clotting time is proportional to the
concentration of factors VIII, IX, XI and XII as well as common pathway factors II, 5. Mix well and incubate at 37 OC for 3 minutes.
V, and X. As the reagent is prepared using one single species rabbit brain, it has 6. Forcibly pipette 100 µL of pre warmed Reagent1 (CaCl2 ) into the test cuvette.
the required sensitivity to be used in heparin assays, also has better sensitivity for
7. Start a timer simultaneously and record the clotting time in seconds.
factors VIII & LA.
Calibration curve for the determination of Heparin concentration:
Kit Components
Dilute Heparin (as used for treatment) with physiological saline to concentration
Reagent/ Product Code Description
of 10 IU/mL.
Component 12602001
Mix 0.2 mL of 10 IU/mL d iluted heparin with 1.8 mL of FNP (Fresh Normal Plasma)
APTT Reagent 1 2 x 4 mL Calcium chloride solution 0.020 M/L to yield heparin standard of 1 IU/mL concentration.
Dilute the Heparin standard as prepared above (1 IU/ml) with FNP as fol lows.
APTT Reagent 2 2 x 4 mL Rabbit brain cephalin
Test Tube 1 2 3 4 5 6 7
Ellagic acid activator
Buffer Heparin std.(1IU/mL) 0.5 0.4 0.3 0.2 0.1 0.1 -
Stabilizers and preservatives FNP in mL - 0.1 0.2 0.3 0.4 0.9 0.5
Heparin con.(IU/mL) 1 0.8 0.6 0.4 0.2 0.1 0
Tri sodium citrate 1 x 10 mL Tri sodium citrate (0.109 M/L) 3.2% Procedure: Manual method to estimate Heparin concentration in the plasma/
sample.
Risk & Safety . Pipette 0.1mL of each Heparin dilution into clean test tubes.
Material Safety data sheets (MSDS) wil l be provided on request. . Add 0.1 mL APTT Reagent (pre-warmed at 37OC). Mix and incubate for exactly
3 minutes at 37OC.
Reagent Storage and Stabil ity . Add. 0.1 mL Pre-warmed Calcium chloride (0.0290 M/L) and simultaneously
The sealed reagents are stable upto the expiry date stated on the label, when stored start stopwatch.
at 2 - 8OC. . Observe clot formation carefully and note the time at the appearance of first
fibrin web.
Precaution . Plot mean of double determination in seconds against each heparin
- Venous blood should be directly drawn into the tube containing anticoagulant. concentration on heparin calibration graph paper provided.
- Ensure that the sample is free of microclots.
. Connect points in a straight line.
- Separate plasma immed iately by centrifugation after col lection of blood.
- The test should be done preferably within two hours of blood col lection. *Plot clotting time of sample on the cal ibration curve and read heparin
- Plasma must be stored in siliconized glass tubes or plastic containers. concentration in IU/mL
- Avoid turbid, lipemic or hemolyzed samples. Note: Laboratories using coagulometers should follow instructions (sequence) of
- Use clean dry micropipette tips and plastic ware to dispense the reagent. the coagulometer manufacturer.
- Mix the reagent (by gentle swirling) before use. Warranty: The product is designed to perform as described in the pack insert. The
- Close reagent vial and replace immediately to 2-8OC after dispensing. manufacturer disclaims any implied warranty of use and sale for any other purpose.

Waste Management Calculation

The result of APTT test can be reported directly in seconds.
Reagents must be disposed off in accordance with local regulations.

Sample Reference Range

Citrated plasma The normal values are between 21-38 seconds (at 3 minutes activation). The normal
time depends on the method, activation time, instrument etc. and must be
Materials provided determined in each laboratory. Results obtained for patient samples are to be
APTT Reagent 1, APTT Reagent 2 and Tri sodium citrate correlated with clinical find ings of patient for interpretation and diagnosis.

Materials Required but Not Provided Bibliography

• Pipettes& Tips 1. Dacie, J.V., Lewis, S.M.; Practical hematology. 1984
• Test Tubes & racks 2. R Biggs, R, Mcfarlane, R. G; Human blood coagulation and its d isorders 1963
• Timer
• Incubator 3. Burtis, et al. Tietz: Text book of Clinical Chemistry AACC 1999
• Analyzer 4. Roadck, B. F; Diagnostic Hematology, clinical principles and applications 2nd
edition.
Sample Col lection & Preparation
5. John Bernad Henry: Clinical Diagnosis and management by laboratory methods
Citrated plasma.
Mix gently, 9 parts of blood in a plastic tube or siliconzied glass tube containing 1 20th edition.
part of 3.2% tri-sod ium citrate solution (0.109 M/L). Centrifuge immediately for 6. Data on file of Agappe Diagnostics Ltd. Kerala.
15 minutes at 3000 rpm to obtain platelet poor plasma. Transfer supernatant plasma
in a siliconized glass tube or plastic tube immediately; do not disturb the buffy coat
while collecting supernatant plasma. The test should be done preferably within 2 hours
of blood collection.
SYMBOLS USED ON THE LABELS

IN VITRO DIAGNOSTIC USE SEE PACKAGE INSERT FOR PROCEDURE LOT NUMBER MANUFACTURER’S ADDRESS MANUFACTURING DATE EXPIRY DATE TEMPERATURE LIMIT

AGAPPE DIAGNOSTICS LTD.

Agappe Hil ls, Dist. Ernakulam, Kerala, India-683 562.
Customer Care No. +91 484 3120002 ISO 9001:2008
[email protected] | www.agappe.com REV. NO.: ADL/IFU/APTT/COA/R00 EN ISO 13485:2012