Page 99 - JSOM Summer 2019

FIGURE 1 Major crossmatch with human red blood cells and canine FIGURE 3 Minor crossmatch with canine DEA 1–positive red
serum. Human blood of various blood types was tested against blood blood cells and human plasma. Red blood cells from five different
samples of 10 canines. Number in parentheses represents number of DEA 1–positive canines were tested against human plasma from
repeated samples. different individuals. Numbers in parentheses represent number of
repeated samples.

FIGURE 2 Minor crossmatch with canine DEA 1–negative red and washed B– RBCs were added to the tubes of heat-treated
blood cells and human plasma. Red blood cells from five different canine plasma. The test tubes were processed and graded as
DEA 1–negative canines were tested against human plasma from described earlier.
different individuals. Numbers in parentheses represent number of
repeated samples.
Statistical Methods
Descriptive statistics were reported as mean ± standard devi-
ation. For the major crossmatch, an omnibus test was used to
determine if significance was present between groups. Pairwise
comparisons were then made between groups with a Bonfer-
2
roni adjustment. For the minor crossmatch, a χ test were used
to analyze categorical variables. Data were analyzed by using
commercial statistical software (Minitab 16, State College,
PA). Significance was set at p < .05.

Results
From the 20 GOC blood samples, blood from 10 healthy
GOCs was used for this study. Seven dogs were German short-
haired pointers, one was a German shepherd, one was a Vizsla,
and one was a Labrador retriever. Seven were intact males,
and three were intact females. Five dogs were DEA 1 positive
than the amount in the original RBC suspensions denoted he- and five were DEA 1 negative. The mean age was 1.5 (± 0.3)
molysis and was recorded as such. Samples without obvious years, and the mean weight was 23.8 (± 3.7) kg. Demographic
macroscopic agglutination were scored microscopically. Two data from the healthy human donors, other than blood type,
drops of the resuspended RBC–serum mixture were placed on were masked to the investigators and could not be reported.
a slide, coverslipped, and evaluated microscopically. No ag-
glutination was scored as “negative.” All samples in this study
showed substantial agglutination and therefore were reported Major Crossmatch
only as agglutination when present. All samples in the major crossmatch showed substantial
cross-reactivity in the form of marked hemolysis or aggluti-
Heat Denaturation Experiment nation. Of the 100 samples tested, 80 showed hemolysis and
20 showed marked agglutination (Figure 1). All eight controls
An additional experiment was conducted immediately after were considered negative for hemolysis or agglutination. A–
the completion of the major and minor crossmatches to try and B– (which showed agglutination) were significantly differ-
to define the role of complement. In this experiment, plasma ent from each other as well as significantly different from the
from four different canines was crossmatched with A–, B–, other blood groups (p < .001 for all). However, only the A–
and O– human RBCs as described here earlier. In addition, and B– samples from the commercial test kit showed aggluti-
100μL of plasma from the same four GOCs was placed into nation (all samples from these test kits agglutinated), whereas
a test tube, capped, and heated in a waterbath at 60°C for 20 all of the samples from individual donors and the O+ test
minutes. These samples were allowed to cool to room tem- kit samples showed marked hemolysis with almost no intact
perature, and then 50μL of A– and O– human RBC suspension RBCs on the slides when evaluated microscopically.

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