D dimer test

Introduction:

D-dimer is a fibrin degradation product(FDP), a

small protein fragment present in the blood after
a blood clot it is degraded by fibrinolysis.
Introduction:

• D-dimer is one of the protein fragments produced

when a blood clot gets dissolved in the body.
• It is normally undetectable or detectable at a very low
level unless the body is forming and breaking down
blood clots.
• Then, its level in the blood can significantly rise. This
test detects D-dimer in the blood
Generation Of D-dimer From Cross-Linked
Fibrin
Reference Range

• D-dimer is the degradation product of cross linked

(by factor XIII) fibrin. It reflects ongoing activation
of the haemostatic system.
• The reference concentration of D-dimer is < 250
ng/mL, or < 0.4 mcg/mL.
Indications

• Elevated level of D-Dimer are found in clinical

conditions such as:
• Deep vein thrombosis
• Pulmonary embolism
• Disseminated intravascular coagulation
Indications

• The D-Dimer level are rise during pregnancy

and high level are associated with complication.
• Normal pregnancy is associated with alterations
of the haemostatic system toward a
hypercoagulable state.
• Elevated markers of coagulation and fibrinolytic
system activation, such as D-dimer, indicate
increased thrombin activity and increased
fibrinolysis following fibrin formation throughout
pregnancy.
Qualitative Method
Material requirement:

• D-dimer latex particle

• Saline solution
• Positive control plasma
• Negative control plasma
• Disposable testing card
• Blood sample
• Pipettes
• Timer
• Mixer rods
Principle :

• D-Dimer proteins in the sample bind to the specific

anti-D-Dimer antibody which is coated with latex
particles, which react with fibrin D-Dimer or
fragment D of fibrin but not with intact fibrinogen. As
a result agglutination occur.
Procedure:

• Bring all the reagents to room temperature.

• Place 20ul of sample in one circle of the test card and
add similar quantity of sample into positive and
negative control.
• Add 20ul of latex reagent into each circle.
• Use separate mixing rods for mixing the contents of
each circle.
• Start the timer.
• Rotate the test card and note the agglutination
between 180-200seconds.
 Procedure:

• Compare the agglutination pattern with those

produced by the positive and negative controls. The
negative control will not give any agglutination but
positive control result will give agglutination.
ICT Method

• Now a days D-dimer is also qualitatively detected

through ICT strip available.
Semi quantitative method:

• This is done only when the qualitative test is positive.

• For this purpose serially dilute 100ul of sample 1:2,
1:4 and 1:8.
• Add 100ul saline solution in these three test tubes.
• Mark the positions of sample dilutions on the test
card and mix with the latex suspension.
• Each dilution is tested similarly.
• The D-dimer concentration may be determined from
the following table:
D-dimer Level (ng/ml)
(ng/ml) Undiluted 1:2 1:4 1:8
<250 - - - -
250-500 + - - -
500-1000 + + - -
1000-2000 + + + -
>2000 + + + +
Interpretation:

• Agglutination occurs within 180-200 seconds for

sample containing greater than 250ng/ml D-dimer.
• The mean level of D-dimer in a healthy population is
between about 8 and 135 ng/ml, and neat plasma
from normal healthy individuals should not give
agglutination.
• The circulatory half-life of D-dimer is about 12
hours.
Quantitative D-dimer Assays

• Quantitative D-dimer assays have replaced semi-

quantitative methods in medical practice. When combined
with clinical criteria, the finding of low D-dimer is useful
for ruling out thrombosis and PTE early in the diagnostic
work-up.
• The Comparative Coagulation Laboratory is now offering
a quantitative D-dimer test for animals in place of a semi-
quantitative test method. A specific value, rather than
concentration range, provides more precise monitoring for
serial assessments of patient status. Testing algorithms
that incorporate quantitative D-dimer may improve
diagnostic accuracy for early identification of thrombosis
Limitations

• False positive readings can be due to various causes:

• Liver disease,
• High Rheumatoid Factor,
• Inflammation,
• Malignancy,
• Trauma,
• Pregnancy,
• Recent surgery.
Limitations
• False Negative readings can occur if the sample is
taken either too early after thrombus formation or if
testing is delayed for several days.
• Additionally, the presence of anti-coagulation can
render the test negative because it prevents thrombus
extension.
References

• Freyburger G, Labrouche S. Comparabilty of D-dimer assays

in clinical samples. Seminars in Vasc Medicine 2005;5:328-
339.
• Arnout J, Sales M, Arza B et al. Clinical management study of
venous thromboembolism using HemosIL D-dimer. (abstr)
ISTH, August 6-12, Sydney Australia, 2005.
• Hart DJ, Hutchman G, Cuthbert RJG. Evaluation of an
automated latex D-dimer immunoassay in the clinical
assessment of suspected venous thromboembolism. Clin. Lab.
Haem. 2002;24:171-174.
• Stokol T, Brooks MB, Erb HN, Mauldin GE. D-dimer
concentrations in healthy dogs and dogs with disseminated
intravascular coagulation Am J Vet Res 2000;61:393-398.