Evaluation of post-transfusion RBC alloimmunization in dogs using a gel-column crossmatch with and without anti-canine globulin enhancement

Abstract

A blood crossmatch is essential to ensure RBC compatibility for previously transfused dogs. There is no gold standard crossmatch method for dogs, although the standards used most commonly by academic institutions and reference laboratories are the tube and gel-column crossmatches. Addition of anti-canine globulin (ACG) has been suggested to increase detection of RBC incompatibilities. Our objective was to determine if there is a correlation between results of a standard and an ACG-enhanced gel-column crossmatch in detecting post-transfusion RBC alloimmunization. Pre- and post-transfusion serum or plasma samples were obtained from 33 dogs for major crossmatches to 1-6 (median: 3) blood donors. Crossmatches were performed with (n = 202) and without (n = 202) ACG, with results scored by 4 observers, 3 of whom were anonymized. Ten of 33 (30%) dogs had major crossmatch incompatibilities post-transfusion. RBC incompatibilities (2-4+ agglutination) were detected only with ACG in 4 dogs, only without ACG in 3 dogs, and with both methods in 3 dogs. There was fair correlation between crossmatch methods for determination of compatibility (ρ = 0.34; p < 0.001) and incompatibility (ρ = 0.35; p < 0.001) scores. Among 4 observers, there was near-perfect agreement in determining compatibility (κ = 0.97; p < 0.001) and substantial agreement in overall scoring of incompatibility (κ = 0.77; p < 0.001). Our results suggest that detection of RBC incompatibilities in dogs can be maximized by performing a gel-column crossmatch both with and without ACG enhancement.

Keywords: alloimmunization; antiglobulin; blood transfusion; crossmatch; dogs.

Figure 1.

Schematic demonstrating the sequence of collection and storage of blood samples from recipient and donor dogs for pre- and post-transfusion crossmatches and auto-controls. CPD = citrate–phosphate–dextrose; pRBC = packed RBC.

Figure 2.

Gel column major crossmatches using pre- and post-transfusion plasma samples (53A and 53B [dog 22], respectively) with RBCs from donor 5243 without (columns 1, 2) and with (columns 3, 4) anti-canine globulin (ACG), demonstrating detection of RBC alloantibodies post-transfusion (3.5+ incompatibility) only with the addition of ACG.