Protime
/Prothrombin(PT)
The prothrombin time is therefore the time
required for the plasma to clot after an excess
of thromboplastin and an optimal
concentration of calcium have been added.
The PT measures functional activity of the
extrinsic and common pathways.
The PT evaluates patients suspected of having
an inherited or acquired deficiency in these
pathways.
PT Reagent
Calcium ions and Thromboplastin from brain tissue
(Rabbit).
Thromboplastin (Tissue Factor) protein-lipid
complex found in tissues outside blood vessels.
Thromboplastins were originally tissue extracts
obtained from different species and different
organs containing tissue factor and phospholipid.
Because of the potential hazard of viral and other
infections from handling human brain, it should no
longer be used as a source of thromboplastin. The
majority of animal thromboplastins now in use are
extracts of rabbit brain or lung.
Principle:
Principle:
The procedure uses a tissue thromboplastin
reagent with CaCl (Calcium Chloride) to provide a
one step procedure for evaluating plasma clotting.
This test was devised on the assumption that
when an optimal amount of calcium and an excess
of thromboplastin are added to decalcified
plasma, the rate of coagulation depends on the
concentration of prothrombin in the plasma.
:Principle
The normal values for the prothrombin time
range from 10.0 to 13.0 seconds.
An elevated prothrombin time may indicate
the presence of
1. Vitamin K deficiency,
2. DIC,
3. liver disease,
4. Presence of FSP’s
:Principle
5. a deficiency in one or more of the following
factors:
1) Factor I (Fibrinogen)
2) Factor II (Prothrombin)
3) Factor V (Proaccelerin, Labile Factor)
4) Factor VII (Proconvertin, Stable Factor)
5) Factor X (Stuart-Prower Factor)
6) Factor XIII (Fibrin Stabilizing Factor)
6. In addition, inhibitors can cause prolonged
PT’s.
When is it
ordered?
Used to monitor oral anticoagulant therapy
(Warfarin / Coumadin).
When a patient who is not taking anti-
coagulant drugs has signs or symptoms of a
bleeding disorder
When a patient is to undergo an invasive
medical procedure, such as surgery, to ensure
normal clotting ability.
Procedure
Reconstitute tissue thromboplastin according to
instructions. Label the thromboplastin with the
time, date and initials. The thromboplastin
reagent is stabile for 7 days after reconstitution.
Allow to sit 10-15 minutes, then invert gently
several times. Mix well prior to pipetting any
of this reagent at any step in this
procedure.
Pipet 1-2 mls, using a plastic pipet, of the tissue
thromboplastin-calcium chloride reagent ( PT
reagent) into a glass test tube and place in a 37
water bath incubator.
Procedure
Pipet 100 µL or .1 mL of normal control, Patient PPP
into each of the test tubes.
Allow at least three minute for the control to reach
37°C.
Pipet 200 µL or .2 mLof PT reagent into the tube
containing the control. Start the stop watch
simultaneously.
Mix the tube and leave in the water bath for a
minimum of 7-8 seconds. Then remove, wipe the
exterior, tilt back and forth gently until a visible clot is
formed. As the clot forms, the mixture will gelatinize
and may turn cloudy.
Procedure
Stop the stop watch immediately when the
clot begins to form and record the time in
seconds. Carry out 1 significant figure passed
the decimal point. For example, if your result
is 12.23 seconds, report as 12.2 seconds.
Repeat the procedure for the second run of
normal control. Record the time.
Procedure
If the results from run 1 and run 2 are within + 1
second from each other, average the two results
and report with appropriate units. For manual PT,
results should match within 1.0 second (if result is
less than 20 seconds). Results over 20 seconds
should match within 2.0 seconds.
If results are not within required limits, a third run
should be performed and average the two that
match within acceptable limits. Be sure and cross
out any values you are not using for the final
calculation. Include measurement unit of
seconds on report sheet.
Interpretation of
Result
The Mean Normal Plasma is used to evaluate routine
result.
The INR is not used to evaluate ROUTINE PT
results.
The International Normalization Ratio (INR) value is
only used on patients who are on oral anticoagulant
therapy such as Coumadin. Because these patients
must be continuously monitored and their dosage
adjusted accordingly, it is very important to
standardize the reported PT results. Standardization
ensures uniformity and safety when a traveling
patient is treated in different geographical areas.
Sources of Error
1. Associated with specimen
a. Inappropropriate ratio of anticoagulant to blood
b. Failure to correct citrate volume if hematocrit > 55%
c. Clotted, hemolyzed or lipemic samples
d. Lack of PPP
e. Delay in testing or processing
f. Inappropriate storage
Sources of Error
1. Associated with storage
a. Incorrect preparation of reagents
b. Failure to properly store reagents
c. Use of reagents beyond reconstituted stability
time or expiration date
d. Contaminated reagents
Sources of Error
1. Associated with procedure
a. Incorrect temperature
b. Incorrect incubation times
c. Incorrect volumes of sample, reagents or both
