Comparison of various blood-typing methods for the feline AB blood group system

OBJECTIVE: To compare feline blood-typing results determined by use of the card (CARD), gel (GEL), tube (TUBE), University of Pennsylvania (Penn) tube, and Penn slide tests. SAMPLE POPULATION: Blood samples from 38 healthy cats. PROCEDURES: Blood samples, anticoagulated with EDTA, were screened by use of each blood-typing method according to manufacturers' protocols. RESULTS: On the basis of the standard Penn tube and slide test results, 20, 11, and 7 cats were classified as type A positive, type B positive, and type AB positive, respectively. The same results were obtained with the anti-B and anti-B reagents of the TUBE test. Use of anti-A antibodies of original polyclonal and current monoclonal CARD tests resulted in mostly 2+ to 3+ (scale, 0 to 4+) agglutination reactions with blood samples from type A-positive cats; agglutination reactions with blood samples from type AB-positive cats were weak (1+). The anti-B lectin of the CARD test induced a 2+ to 4+ reaction with blood from all type B- and type AB-positive cats. Use of the GEL test allowed recognition of type A and type B blood samples; following addition of anti-A serum to control columns, type B blood was differentiated from type AB blood. CONCLUSIONS AND CLINICAL RELEVANCE: Use of the in-practice CARD test allows identification of type A- and type B-positive cats, but weak reactions of type AB blood with the anti-A monoclonal antibody raise concerns. The modified GEL and TUBE tests appear to be reliable clinical laboratory methods for feline blood typing.     

Comparison of five blood-typing methods for the feline AB blood group system

Objective-To compare the ease of use and accuracy of 5 feline AB blood-typing methods: card agglutination (CARD), immunochromatographic cartridge (CHROM), gel-based (GEL), and conventional slide (SLIDE) and tube (TUBE) agglutination assays. Sample Population-490 anticoagulated blood samples from sick and healthy cats submitted to the Transfusion or Clinical Laboratory at the Veterinary Hospital of the University of Pennsylvania. Procedures-Sample selection was purposely biased toward those from anemic, type B, or type AB cats or those with autoagglutination. All blood samples were tested by use of GEL, SLIDE, and TUBE methods. Fifty-eight samples were also tested by use of CARD and CHROM methods. The presence of alloantibodies in all cats expressing the B antigen as detected by use of any method was also assessed. Results-Compared with the historical gold-standard TUBE method, good to excellent agreement was achieved with the other typing tests: CARD, 53 of 58 (91% agreement); CHROM, 55 of 58 (95%); GEL, 487 of 490 (99%); and SLIDE, 482 of 487 (99%; 3 samples were excluded because of autoagglutination). Four of the samples with discordant test results originated from cats with FeLV-related anemia. Conclusions and Clinical Relevance-Current laboratory and in-clinic methods provide simple and accurate typing for the feline AB blood group system with few discrepancies. Retyping after in-clinic typing with the GEL or TUBE laboratory methods is recommended to confirm any type B or AB cats.     

Dog erythrocyte antigens 1.1, 1.2, 3, 4, 7, and Dal blood typing and cross‐matching by gel column technique

Background: Testing for canine blood types other than dog erythrocyte antigen 1.1 (DEA 1.1) is controversial and complicated by reagent availability and methodology. Objectives: The objectives of this study were to use available gel column technology to develop an extended blood‐typing method using polyclonal reagents for DEA 1.1, 1.2, 3, 4, 7, and Dal and to assess the use of gel columns for cross‐matching. Methods: Dogs (43–75) were typed for DEA 1.1, 1.2, 3, 4, 7, and Dal. Methods included tube agglutination (Tube) using polyclonal reagents, a commercially available DEA 1.1 gel column test kit (Standard‐Gel) using monoclonal reagent, and multiple gel columns (Extended‐Gel) using polyclonal reagents. Blood from 10 recipient and 15 donor dogs was typed as described above and cross‐matched using the gel column technique. Results: Of 43 dogs typed for DEA 1.1, 23, 25, and 20 dogs were positive using Standard‐Gel, Extended‐Gel, and Tube, respectively. Typing for DEA 1.2 was not achievable with Extended‐Gel. For 75 dogs typed for DEA 3, 4, and 7, concordance of Extended‐Gel with Tube was 94.7%, 100%, and 84%, respectively. Dal, determined only by Extended‐Gel, was positive for all dogs. Post‐transfusion major cross‐matches were incompatible in 10 of 14 pairings, but none were associated with demonstrable blood type incompatibilities. Conclusions: Gel column methodology can be adapted for use with polyclonal reagents for detecting DEA 1.1, 3, 4, 7, and Dal. Agglutination reactions are similar between Extended‐Gel and Tube, but are more easily interpreted with Extended‐Gel. When using gel columns for cross‐matching, incompatible blood cross‐matches can be detected following sensitization by transfusion, although in this study incompatibilities associated with any tested DEA or Dal antigens were not found.     

Production, characterization, and applications of a murine monoclonal antibody to dog erythrocyte antigen 1.1.

A murine IgM monoclonal antibody, which recognizes dog erythrocyte antigen (DEA) 1.1, has been produced. The antibody correctly identified canine RBC possessing DEA 1.1 in a panel of RBC typed by an independent laboratory. Reactivity of the monoclonal antibody was compared with canine anti-DEA 1.1 antiserum with 163 RBC samples from 145 dogs. Results of agglutination tests with the 2 reagents were in agreement for all samples. A card agglutination test that uses the monoclonal antibody with blood is described. A monoclonal antibody-based test should facilitate blood typing for DEA 1.1 in clinical practice.     

Prevalence of dog erythrocyte antigens in retired racing Greyhounds

Background: Blood groups in dogs are designated as dog erythrocyte antigen (DEA) 1.1, 1.2, 3, 4, 5, 7, and Dal. There is limited information about the frequency of different antigens in Greyhound dogs, despite their frequent use as blood donors. Objectives: The aims of this study were to determine the frequencies of DEA 1.1, 1.2, 3, 4, 5, and 7 in Greyhounds, to compare the frequencies with those of non‐Greyhound dogs, and to evaluate the presence of naturally occurring anti‐DEA antibodies. Methods: Blood was collected from 206 Greyhound and 66 non‐Greyhound dogs being screened as potential blood donors. Blood‐typing was performed at Animal Blood Resources International by tube agglutination utilizing polyclonal anti‐DEA antibodies. Results: Of the Greyhound dogs, 27/206 (13.1%) were positive for DEA 1.1, and this frequency was significantly lower (P<.0001) than for non‐Greyhound dogs of which 40/66 (60.6%) were DEA 1.1‐positive. The frequency of positivity for both DEA 1.1 and 1.2 was also lower in Greyhounds (P<.0001). There were no significant differences between Greyhounds and non‐Greyhounds for DEA 1.2, 3, 4, 5, or 7. All 137 dogs (113 Greyhounds and 24 non‐Greyhounds) that were evaluated for naturally occurring anti‐DEA antibodies in serum were negative. A higher percentage of Greyhound dogs (57.3%, 118/206) were considered “universal donors” (negative for all DEAs except DEA 4) compared with non‐Greyhound dogs (28%, 13/46). Conclusion: The frequency of positivity for DEA 1.1 in our population of Greyhounds was significantly lower than previously reported for dogs. Furthermore, a large majority of Greyhounds met the criteria for universal donors.     

Canine Dal blood type: A red cell antigen lacking in some Dalmatians

BACKGROUND: Based upon alloantibodies produced after sensitizing dogs with transfused blood, more than a dozen blood group systems have been recognized thus far, and some have been classified as dog erythrocyte antigens (DEA). HYPOTHESIS: A new canine red cell antigen was suspected, based on the development of specific alloantibodies in a Dalmatian previously sensitized by blood transfusions. ANIMALS: Twenty-six Dalmatians (including 1 Dalmatian in need of blood compatibility studies); 55 canine blood donors. METHODS: Serologic tests, including blood typing, crossmatching, and direct Coombs' test were performed by standard tube techniques and a novel gel column technology adapted from human blood banking. RESULTS: By day 40 after transfusion of an anemic Dalmatian, all major crossmatch tests to 55 non-Dalmatian dogs were incompatible. The 2 initial donors, who were compatible before transfusion, were also now incompatible, suggesting the development of an alloantibody to a common red cell antigen. No siblings were available, but 4 of 25 unrelated Dalmatians were crossmatch compatible, suggesting that they were missing the same red cell antigen. The patient was blood typed DEA 1.1, 3, 4, and 5 positive, but DEA 7 negative. Further blood typing and crossmatching results did not support an association to any of these known blood types. The alloantibodies produced were determined to be of the immunoglobulin G class. CONCLUSIONS AND CLINICAL IMPORTANCE: Based upon the identification of an acquired alloantibody in a Dalmatian, a presumably new common blood type named Dal was identified. Dalmatians lacking the Dal antigen are likely at risk of delayed and acute hemolytic transfusion reactions.     

Hemolytic Transfusion Reactions in a Dog With an Alloantibody to a Common Antigen

Alloantibodies to high-frequency red cell antigens, defined as inherited traits occurring in 92% to 99% or more of the general population, are recognized as a cause of hemolytic transfusion reactions in humans. Here we describe a dog (dog erythrocyte antigen [DEA] 1.2-and DEA 4-positive) sensitized by prior blood transfusion, for which a compatible blood donor could not be found; transfusion of DEA 1.1-negative blood resulted in hemolytic transfusion reactions. Patient serum from days 1 (before first transfusion) and 16 was available for further testing; using 4 dogs with different blood types as potential donors, the major crossmatches were compatible using serum from day 1. However the crossmatches were all incompatible with serum from day 16, indicating that the patient was sensitized to an antigen after the first transfusion. The presence of an alloantibody against DEA 1.1 was not ruled out in this patient, but the incompatibility reactions of patient serum with red cells from donors negative for DEA 1.1 indicated that an alloantibody against a red cell antigen other than DEA 1.1 or any other known DEA for which typing reagents were available (DEA 3, 5, and 7) was present. Subsequently, red cells from 1 of the patient's siblings (DEA 1.2-, 4-, and 7-positive) were found not to agglutinate when incubated with patient's serum from day 16, ruling out the presence of an anti-DEA 7 antibody, and suggesting that an alloantibody against a common red cell antigen missing in the patient and sibling was responsible for the blood incompatibility reactions. Failure to obtain a compatible crossmatch with several universal donors in a dog previously transfused should raise a suspicion that an alloantibody to a common red cell antigen may exist and that a sibling may be a source of compatible blood.     

The breed prevalence of dog erythrocyte antigen 1.1 in the Onderstepoort area of South Africa and its significance in selection of canine blood donors

The blood group antigen Dog Erythrocyte Antigen (DEA) 1.1 is clinically the most important canine blood group as DEA 1.1 antibodies are capable of causing acute haemolytic, potentially life-threatening transfusion reactions. Dogs do not have naturally occurring antibodies to DEA 1.1 but are rapidly sensitised by the first incompatible transfusion. The prevalence of DEA 1.1 in the general dog population is estimated at 42-46%. Canine blood donors registered with the Onderstepoort Animal Blood Bank (n = 93) as well as potential donors (n = 140) were typed for DEA 1.1 using a monoclonal antibody card kit. All dogs came from the Onderstepoort area, near Pretoria, Gauteng province, South Africa. Overall prevalence of DEA 1.1 was 47%. Prevalence was 47% in purebred dogs and 48% in mongrels. Distinct breed differences were noted with less than 20% of German shepherd dogs and Boxers and greater than 75% of Rottweilers, Great Danes, St Bernards and Dalmations testing DEA 1.1 positive. Knowledge of local breed differences will increase effectiveness of blood donor recruitment.     

Prevalence of dog erythrocyte antigen 1.1 in dogs in Switzerland evaluated with the gel column technique

Canine blood typing has become an established and essential laboratory test due to the rising demand for safe and efficient blood transfusions. The most immunogenic and clinically important blood type is DEA 1.1. Little is known about DEA 1.1 frequencies or special characteristics among different canine breeds. 304 dogs were tested for DEA 1.1. DEA 1.1-typing was performed using a commercial gel column technique (ID-Gel Test Canine DEA 1.1, DiaMed, Cressier, Switzerland). Fifty-three percent of all tested dogs reacted positive for DEA 1.1, whereas 49 % of the mixed breeds tested DEA 1.1-positive. All Bernese mountain dogs (n = 22) and Rottweilers (n = 9) tested positive for DEA 1.1, while all Boxers (n = 8), Flat-Coated Retrievers (n = 9), and Border Collies (6) tested negative for DEA 1.1. The prevalence of DEA 1.1 in dogs in Switzerland was found to be comparable to that reported from other countries. The tested breeds were found to differ considerably in the frequency of DEA 1.1. This knowledge is useful for selection of blood donors. However, DEA 1.1 blood typing of donor and recipient prior to transfusion and cross matching in sensitized dogs is unavoidable.     

Pattern of Coombs' test reactivity has diagnostic significance in dogs with immune-mediated haemolytic anaemia

OBJECTIVES: To investigate the clinical significance of the pattern of Coombs' test reactivity in dogs with immune-mediated haemolytic anaemia. METHODS: Sixty-five anaemic dogs with a positive Coombs' test were included. Coombs' testing was performed at 4 and 37 degrees C with polyvalent canine Coombs' reagent and antisera specific for each of canine immunoglobulin G, immunoglobulin M and complement factor C3. The impact of performing testing with only polyvalent antiserum at 37 degrees C was assessed. Chi-squared tests were used to compare Coombs' test reactivity in dogs with primary immune-mediated haemolytic anaemia (group A) and in dogs with concurrent/underlying disease (group B). Following Bonferroni correction, significance was set at P < or = 0.003. RESULTS: Eleven dogs would have been regarded as Coombs' negative had they been tested with polyvalent antiserum at 37 degrees C alone. Group A dogs were significantly more likely to be positive with polyvalent antiserum and/or anti-dog immunoglobulin G at 4 and/or 37 degrees C (P < or = 0.001) and tended to be less likely to be positive with anti-dog immunoglobulin M at 4 degrees C (P=0.040). CLINICAL SIGNIFICANCE: Testing of anaemic dogs with polyvalent Coombs' reagent at 37 degrees C was less sensitive than testing with monovalent reagents at 4 and 37 degrees C. The pattern of Coombs' test reactivity differed significantly between dogs with primary immune-mediated haemolytic anaemia and those with concurrent/underlying disease.     

A comparison of standard serological tests for the diagnosis of bovine brucellosis in Canada.

Six agglutination and two complement fixation tests were compared with respect to specificity, sensitivity and relative sensitivity for the serodiagnosis of bovine brucellosis. Based on 1051 sera from brucellosis free herds, the specificity of the tests was 98.9% for the buffered plate antigen test (BPAT), 99.2% and 99.3% for the standard tube and plate agglutination tests (STAT and SPAT), respectively, and 99.8% for the 2-mercaptoethanol test (2MET). On this small sample, the rose bengal plate test (RBPT), card test (CARD) and the complement fixation test (CFT) correctly classed all sera as negative. On a sample of 167 culture positive cattle, the sensitivities of the tests were CFT: 79.0%, BPAT: 75.4, RBPT: 74.9%, CARD: 74.3%, SPAT: 73.1%, STAT: 68.9%, and 2MET: 59.9%. All tests combined detected only 82% of these infected cattle. Analysis of the relative sensitivity of the six agglutination tests gave the following ranking: BPAT greater than RBPT greater than CARD greater than SPAT greater than STAT. The 2MET ranked between the BPAT and RBPT or between the RBPT and CARD depending on the analysis used. The use of the BPAT as a screening test is recommended provided that a test of high specificity and sensitivity such as the CFT is used to confirm screening test reactions.     

Lack of Evidence of Pregnancy-Induced Alloantibodies in Dogs

Background: It is controversial whether or not pregnant bitches become sensitized to red blood cell (RBC) antigens.
Hypothesis: Bitches do not develop alloantibodies to RBC antigens during gestation and can be used safely as blood donors.
Animals: The study group included 35 healthy female dogs with a prior history of 1 (n = 12), 2 (n = 14), or ≥ 3 (n = 9) pregnancies. The control group consisted of 15 healthy female dogs without any history of pregnancy.
Methods: All dogs were blood typed for dog erythrocyte antigens (DEA) 1.1, 1.2, 3, 4, 5, and 7 using ethylenediaminetetraacetic acid blood samples and polyclonal antisera. Antibody screening was performed with serum and canine RBC panels of known blood type. An autocontrol and direct antiglobulin test were performed to rule out the presence of autoantibodies.
Results: The only alloantibodies identified were those against DEA 7 and the prevalence of anti-DEA 7 alloantibodies was similar in dogs with known history of pregnancy (11.4%) and in the control group (13.3%).
Conclusions and Clinical Importance: These results confirm previous studies and clinical transfusion medicine experience. Naturally occurring anti-DEA 7 alloantibodies have been reported but their clinical relevance has not been shown. Pregnancy does not appear to sensitize dogs to RBC antigens. Consequently, dogs with prior history of pregnancy can be used safely as blood donors. Conversely, no additional pretransfusion compatibility studies would be required should these dogs themselves need to be transfused.     

Characterization of a new fimbrial antigen present in Escherichia coli strains isolated from calves.

Thirteen Escherichia coli strains isolated from calves with diarrhoea, supposed to carry a common antigen were examined for their hemagglutinating activity and compared by bacterial agglutination, double diffusion in two dimensions and by crossed immunoelectrophoresis (CIE). Two of the strains were examined also in the electron microscope. Most of the strains agglutinated red blood cells of horse, ox, guinea pig and chicken, of which the agglutination of ox erythrocytes was mannose-resistant (MRHA). None of the strains agglutinated human erythrocytes. All strains with MRHA of ox red blood cells, regardless to their O:K:H antigens could be agglutinated in unabsorbed or absorbed antisera produced against cultures C1209 (020:K-:H9) and C1213 (09:K36:H-) when live cells as antigens were used. None of these sera agglutinated reference strains carrying K88, K99, 987P, F41 or FY (Att25) antigen respectively. By the double gel diffusion test and by CIE in extracts (60 degrees C) of the strains a common heat labile antigen, responsible for the MRHA of ox red blood cells was identified. Electron microscopy revealed that this common antigen was represented by thin, long, hair-like fimbriae on cells of E. coli C1213, and that specific homologous antibodies attached to these fimbriae.     

Evaluation of a rapid agglutination method for detection of equine red cell surface antigens (Ca and Aa) as part of pretransfusion testing

BACKGROUND: Blood typing before transfusion minimizes the risk of transfusion reactions and prevents immunization of the recipient against incompatible RBC antigens. The major RBC antigens that warrant identification before packed RBC or whole blood transfusions in horses are Ca and Aa. Standard blood-typing protocols are time-consuming (2.5-3.0 hours) and impractical in emergency settings. OBJECTIVES: The purpose of this study was to determine whether equine RBCs could be typed for Ca and Aa antigens using sera from horses with RBC antibodies in a modified rapid (15 minute) blood-typing protocol. METHODS: Serum was obtained from a horse with anti-Ca antibodies and from another horse with anti-Aa antibodies. The presence of agglutinating antibodies was confirmed with antibody screening. Venous blood samples, collected in citrate-phosphate-dextrose, were obtained from 21 horses of various breeds. Samples were blood typed in the Veterinary Medical Teaching Hospital Hematology Laboratory using standard methodology. Washed RBCs from each of the 21 horses were incubated individually with anti-Ca and anti-Aa sera at dilutions of 1:4, 1:8, and 1:16 for 15 and 30 minutes at room temperature and 37 degrees C. RESULTS: Of the 21 horses, 13 were identified as Aa+/Ca+, four were Aa+/Ca-, two were Aa-/Ca+, and two were Aa-/Ca-. All 17 Aa-positive horses had a positive agglutination reaction at all dilutions of anti-Aa serum, incubation times, and temperatures, while all Aa-negative horses were negative. Each Ca-positive horse had a positive agglutination reaction at all incubation time points and temperatures up to the 1:16 dilution of the anti-Ca serum. All Ca-negative horses were negative at all times, temperatures, and dilutions of anti-Ca serum. Use of the modified protocol on 26 hospitalized horses resulted in accurate typing, based on complete antibody screens. CONCLUSIONS: These results support the hypothesis that equine RBCs can be blood typed using a rapid (15 minute) protocol, at room temperature, for the presence of Ca and Aa antigens using equine-derived antisera. This technique may be beneficial for pretransfusion testing of equine patients in an emergency setting.     

Effect of immune-mediated erythrocyte agglutination on analysis of canine blood using a multichannel blood cell counting system

Blood from six dogs with in vitro immune-mediated erythrocyte agglutination resulted in analytical errors in directly measured counting and sizing functions on a multichannel blood analysis system with histogram capability. Errors in the directly measured values, mean cell volume (MCV), and erythrocyte count were attributed to agglutinated erythrocyte particles that persisted during the relatively short reagent contact time of the analysis. Agglutinated particles less than 240 fl were visible on erythrocyte histograms and resulted in a false low erythrocyte count and false high MCV. Agglutinated cell particles greater than 240 fl were not present on the histogram scale. Because these latter particles exceeded the upper threshold, they did not influence determination of MCV, but resulted in a further decrease in the erythrocyte count. As a result, the other dependent erythrocyte indices were in error. These included false low hematocrit and false high mean corpuscular hemoglobin concentration (MCHC), when compared to corrected reference blood values. Similar errors occurred when analyzing blood samples that were agglutinated in vitro by incubating erythrocytes with incompatible plasma. The counting and sizing errors observed with electronic counting techniques were eliminated or greatly reduced by incubating blood in cell counting diluent for 10 minutes followed by analysis on a single channel counter with attached particle size analyzer. Error in erythrocyte measurement on a multichannel system may be anticipated if there is overt erythrocyte agglutination in a blood sample, an abnormally high MCHC is reported by the system, or subpopulations of large volume (agglutinated cells) are observed on a volume distribution histogram.     

Detection of von Willebrand's Disease in Dogs with a Rapid Qualitative Test, Based on Venom-Coagglutinin-Induced Platelet Agglutination

A rapid, qualitative blood test was developed to identify potential bleeders among dogs that are von Willebrand's disease suspects. The test, based on venom coagglutinin-induced agglutination of formalin-fixed washed platelets, was compared to the more lengthy electroimmunoassay for factor VIII-related antigen. The 2 tests were performed on plasma samples from 82 dogs, including 68 Doberman pinschers. The 39 plasma samples that were found to be "clinically normal" by the new test had factor VIII-related antigen concentrations ranging from 32% to 192% of that in a pool of normal canine plasma samples. Plasma samples from the 43 dogs that tested as "potential bleeders" by the new test had factor VIII-related antigen concentrations from 4% to 38%. All 43 "potential bleeders" were Doberman pinschers. Shortly after being evaluated by the new test, 46 of the dogs underwent invasive diagnostic, therapeutic, or cosmetic procedures. Excessive hemorrhage was encountered in 40% of the 25 dogs that tested as "potential bleeders. " In contrast, only 5% of the 21 dogs that tested as "clinically normal" exhibited abnormal bleeding.     

An investigation of the role of immunologic factors in anemia associated with canine heartworm disease

Direct Coombs' antiglobulin tests were performed on 80 dogs with patent Dirofilaria immitis infection and 170 dogs negative for microfilaria of D. immitis. Presence or absence of anemia was determined by hematocrit in 55 of the heartworm negative dogs and 68 of the dogs with heartworm disease. Heartworm infected dogs showed a higher incidence (37%) of anemia than noninfected dogs (14.5%). Anemia was most prevalent in two groups of dogs with heartworm infection, one group showing vena caval syndrome (91%) and the other occult dirofilariasis (62.5%). These latter two groups of dogs also showed a significantly higher number of positive Coombs' reactions at 37 degrees C than other dogs with heartworm disease and the noninfected dogs. The number of positive Coombs' reactions at 4 degrees C among the total of 80 dogs with heartworm infection was significantly higher than that for dogs without heartworm disease. However, there was no positive correlation between anemia and the outcome of the Coombs' test at either temperature. These findings do not suggest that immunologic factors play a major role in the pathogenesis of anemia in dogs with heartworm disease.     

Some serological reactions to "brucella" antigen in the horse.

Seventy-three samples of serum, from 69 horses and one zebra, were subjected to the Rose Bengal Plate, serum tube agglutination, complement fixation, and anti-equine globulin (Coombs') tests for brucellosis. Fifty-one of the samples, from 48 horses, were submitted by practising veterinary surgeons; of these, 22 samples were associated with clinical conditions which might have been due to brucellosis. Fourteen samples were from healthy horses known to have been in contact with infected cattle, and six were from horses which were known not to have been exposed to brucellosis. More reactions at accepted diagnostic levels were obtained to the anti-globulin and complement fixation tests than to the agglutination test. Two horses showed no reactions, other than inconclusive titres to the antiglobulin test, and these titres could have resulted from exposure to Brucella. Eight of nine positive Rose Bengal tests were confirmed by a reaction at diagnostic level in at least one of the other tests, but two sera showing a reaction at diagnostic level to the other tests gave a negative Rose Bengal result.     

Case-control study of blood type, breed, sex, and bacteremia in dogs with immune-mediated hemolytic anemia

OBJECTIVE: To determine whether blood type, breed, or sex were risk factors for immune-mediated hemolytic anemia (IMHA) in dogs and whether bacteremia was common in dogs with IMHA. DESIGN: Case-control study. ANIMALS: 33 dogs with IMHA, 1,014 dogs without IMHA for which blood type (dog erythrocyte antigens 1.1, 1.2, 3, 4, 5, and 7) was known, 15,668 dogs without IMHA for which breed was known, and 15,589 dogs without IMHA for which sex was known. PROCEDURE: Blood type, breed, and sex distribution of dogs with IMHA were compared with data for control dogs with Fisher exact tests and by calculating odds ratios (ORs). Results of bacterial culture of blood samples were documented for dogs with IMHA, when available. RESULTS: Dog erythrocyte antigen 7 was associated with a significant protective effect (OR, 0.1) in Cocker Spaniels with IMHA (n = 10), compared with control dogs. Cocker Spaniels, Bichon Frise, Miniature Pinschers, Rough-coated Collies, and Finnish Spitz had a significantly increased risk of IMHA, as did female dogs (OR, 2.1). Blood samples from 12 dogs with IMHA were submitted for bacterial culture, and none had bacteremia. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that blood type, breed, and sex may play a role in IMHA in dogs.