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The improvement of gel coombs card
The Direct Coombs/Antiglobulin test (DCT)
was originally described by Coombs, Moorant and Race in 1945. The principle of
the test is to demonstrate antibodies or complement coating red cells invivo by
using Antihuman globulin (AHG) or Coombs reagent.
Till date it remains the hallmark for the
diagnosis of immune hemolytic anemias. Technically various modifications in the
procedure have been described to bring about added sensitivity to the DCT. Some
of these changes include: use of more specific reagents like monospecific AHGs
(e.g. against IgG or C3) and Enzyme elution of the antibodies from the red
cells. In practice, it was noted that DCT required technical expertise (esp
steps of washing the red cells, effective centrifugation etc.) for proper
interpretation.
In 1990 Lappierre introduced the novel ‘GEL’ method to
demonstrate antibodies or complement coated red cells. The gel system is based
on the principle that the Sephadex gel matrix acts as a sieve, through which
agglutinates of RBCs are too large to pass though and remain entrapped in the
gel, depending on their size. A negative reaction is seen as a clear pellet of
cells settled at the bottom of the microtube. Grading of the reactions can be
done according to the distribution of RBCs agglutinates through the gel column.
This technique was easier to perform and did not require technical skill,
thereby overcoming the practical difficulties of performing DCTs.
In this context we have tried to evaluate
three of the methods used for the interpretation of DCT gel coombs card.
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