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 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.