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1.
S. Tasker J. K. Murray T. G. Knowles M. J. Day 《The Journal of small animal practice》2010,51(4):192-199
Objective : To investigate the associations between Coombs’ testing, haemoplasma and retroviral infections, and feline anaemia. Methods : Haematology, Coombs’ testing (including assessment of persistent autoagglutination) and selected infection testing (haemoplasma, feline leukaemia virus/feline immunodeficiency virus provirus) were performed in blood samples collected from 60 anaemic and 60 non-anaemic cats. Results : No association between infection and anaemia or Coombs’ positivity existed. Anaemic cats (21.7%) were significantly more likely than non-anaemic cats (0%) to have cold autoagglutination (P<0.0001), but significance (set at ≤0.0025 due to multiple testing) was not quite reached when Coombs’ positivity was compared between anaemic (40.4% and 21.7% positive at 4°C and 37°C, respectively) and non-anaemic (20% and 3.3% positive, P=0.021 and P=0.004, at 4°C and 37°C, respectively) cats. Cats with immune-mediated haemolytic anaemia were significantly more likely to have persistent cold autoagglutination (P<0.0001) and be Coombs’ positive at 37°C with polyvalent (P<0.0001), immunoglobulin (Ig)G (P<0.0001) or any antiserum (P<0.0001). Haemoplasmas and retroviruses were uncommonly detected. Clinical Significance : Cats suspected of having immune-mediated haemolytic anaemia should be evaluated for persistent autoagglutination at 4°C as well as performing Coombs’ testing at 37°C, but positive results may occur in with other forms of anaemia. Testing for erythrocyte-bound antibodies should always be interpreted in parallel with documentation of haemolysis in anaemic cats. 相似文献
2.
Antiglobulin (Coombs') reagents were assessed for their ability to detect immunoglobulin and complement attached to red cells. Polyspecific and monospecific reagents were prepared using a number of immunisation protocols. Performance of these antisera against control red cells was compared, in a direct Coombs' test, with samples from cases of canine autoimmune haemolytic anaemia (AIHA). A combined reagent containing two monospecific antisera (anti-IgG + anti-C3) gave optimum results. Positive control red cells were required to standardise canine Coombs' reagents for the laboratory diagnosis of AIHA. The optimum incubation temperature for the canine Coombs' test was shown to be 37 degrees C. 相似文献
3.
Florence Juvet Urs Giger Ian Battersby Pierre Menaut Harriet M Syme Carmel T Mooney 《Irish veterinary journal》2013,66(1):12
Erythrocyte pyruvate kinase (PK) deficiency is described for the first time in three apparently unrelated West Highland white terriers (WHWT) from Ireland and the UK. All three dogs were diagnosed with markedly regenerative but persistent anaemia and had been treated for presumed immune-mediated haemolytic anaemia (IMHA) before hereditary erythrocyte PK-deficiency was confirmed by breed-specific DNA mutation analysis. This hereditary erythroenzymopathy causes haemolytic anaemia and affects several canine breeds with varying degrees of severity. Although eventually causing osteosclerosis, haemosiderosis and death, PK-deficient dogs can adapt to their anaemia for many years.PK-deficiency should be considered in anaemic WHWTs worldwide particularly in dogs with haemolytic anaemia where evidence for an immune-mediated, infectious or toxic underlying cause is lacking. 相似文献
4.
Foster AP Sturgess CP Gould DJ Iwasaki T Day MJ 《The Journal of small animal practice》2000,41(6):266-270
A seven-year-old neutered male cocker spaniel was presented with an 11-month history of generalised bacterial dermatitis. There were skin lesions over the entire body, which were round, slightly raised and encrusted. Skin biopsies were collected and the histological findings were consistent with pemphigus foliaceus. Immunohistochemical staining by the indirect immunoperoxidase method was positive, with desmosomal deposition of immunoglobulin (Ig) G. Haematological analysis revealed a regenerative anaemia and profound thrombocytopenia, while a Coombs' test was positive for polyvalent canine Coombs' reagent and anti-dog IgG. An antinuclear antibody test was positive, with a titre of 10,240. An ophthalmic examination demonstrated low tear production (keratoconjunctivitis sicca). Seven months after initial referral, the dog was re-presented with severe generalised peripheral lymphadenopathy. Radiographic evaluation of the thorax and abdomen revealed enlarged cranial mediastinal and sublumbar lymph nodes. Tru-Cut biopsy from an enlarged lymph node confirmed the diagnosis of lymphoma, which was phenotyped as a B-cell tumour. The diagnosis in this case was systemic lupus erythematosus, with the unusual feature of pemphigus foliaceus, and subsequent development of B-cell lymphoma. The case adds further to knowledge of the protean clinical presentations of canine autoimmune diseases and provides additional evidence for the potential association between autoimmunity and immune-system neoplasia in this species. 相似文献
5.
Overmann JA Sharkey LC Weiss DJ Borjesson DL 《Veterinary clinical pathology / American Society for Veterinary Clinical Pathology》2007,36(2):179-183
BACKGROUND: The Coombs' test, which detects immunoglobulin or complement on RBC surfaces, has long been the standard for laboratory confirmation of immune-mediated hemolytic anemia (IMHA), a common cause of hemolytic anemia in the dog. This test, however, suffers from relatively low sensitivity. Optimization of test sensitivity would lead to fewer discrepancies between laboratory results and clinical diagnoses, and in some cases institution of appropriate therapy in a timely manner. OBJECTIVES: The objectives of this study were to 1) characterize the sensitivity and specificity of 2 canine Coombs' tests for the detection of IMHA, 2) document the efficacy of using multiple antisera dilutions beyond what is directed by manufacturers, and 3) evaluate the necessity of monovalent antisera in the test protocol. METHODS: Sixty-five canine whole-blood samples submitted for Coombs' testing were evaluated. Patients were classified as IMHA positive or negative based on a set of predetermined criteria. IMHA classification was compared to Coombs' test results from 2 different Coombs' tests adapted to a microtiter-plate format. One Coombs' test (VMRD Coombs' test) utilized a single polyvalent antiserum (VMRD, Inc, Pullman, WA, USA), while a second Coombs' test (University of Minnesota [U of MN] Coombs' test) used both polyvalent and monovalent antisera. RESULTS: Sensitivity and specificity were 61% and 100% for the VMRD Coombs' test, and 82% and 95% for the U of MN Coombs' test. The use of multiple antisera dilutions resulted in 6 additional Coombs' positive test results. All positive Coombs' test results were positive by polyvalent antisera. CONCLUSIONS: When used in a microtiter-plate format, the U of MN Coombs' test was a more sensitive test for the detection of IMHA in canine patients when compared to the VMRD Coombs' test. The use of multiple antisera dilutions increased test sensitivity. Sensitivity, however, was not increased by the use of monovalent antisera in the Coombs' test protocol. 相似文献
6.
Objectives : To establish whether heightened platelet activation is a common feature of canine immune-mediated haemolytic anaemia, and to evaluate the hypothesis that platelet activation plays a role in the pathogenesis of thromboembolism. Methods : Using whole-blood flow-cytometric analysis, the proportion of activated platelets and platelet-leucocyte aggregates in blood samples from 14 dogs with immune-mediated haemolytic anaemia and 14 healthy dogs was calculated. General linear models with binomial errors were used to compare groups. Results from the immune-mediated haemolytic anaemia-affected dogs were then correlated with established risk factors for thromboembolism in canine immune-mediated haemolytic anaemia, D-dimer concentration and antithrombin activity. Results : There was a strong correlation between platelet activation and severe thrombocytopenia, with heightened platelet activation being observed predominantly in severely thrombocytopenic dogs. Clinical Significance : Dogs with immune-mediated haemolytic anaemia, particularly those with concurrent severe thrombocytopenia, are likely to have heightened platelet activation, which may play a role in the pathogenesis of thromboembolism. 相似文献
7.
Warman SM Helps CR Barker EN Day S Sturgess K Day MJ Tasker S 《The Journal of small animal practice》2010,51(10):534-539
Objective : The aim of this study was to investigate whether the two canine haemoplasma species, Mycoplasma haemocanis and “Candidatus Mycoplasma haematoparvum,” are commonly associated with immune-mediated haemolytic anaemia (IMHA) in UK dogs. Methods : Three groups of dogs were recruited to the study: anaemic dogs with primary IMHA (n=37); anaemic dogs not meeting the inclusion criteria for primary IMHA (n=77) and non-anaemic dogs (n=113). DNA was extracted from 100 μl of blood and subjected to real-time quantitative polymerase chain reaction (qPCR) assays for both species of Mycoplasma. Each assay incorporated co-amplification of canine glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as an endogenous internal control. Results : Canine GAPDH was successfully amplified by qPCR from all 227 canine blood samples but none contained M. haemocanis or “Candidatus M. haematoparvum” DNA. Clinical Significance : Haemoplasma infection is uncommon in dogs in the UK and no evidence was found that these organisms act as triggers for IMHA. 相似文献
8.
Borreliosis in dogs from southern Connecticut 总被引：13，自引：0，他引：13
L A Magnarelli J F Anderson A F Kaufmann L L Lieberman G D Whitney 《Journal of the American Veterinary Medical Association》1985,186(9):955-959
Blood samples were obtained from dogs in tick-infested regions of southern Connecticut to assess canine exposure to Borrelia burgdorferi, the etiologic agent of Lyme disease in human beings. An indirect fluorescent antibody test detected immunoglobulin (Ig)M antibodies at titers of 1:64 to 1:512 in 22 of 84 serum samples previously shown to be positive with a polyvalent rabbit anti-dog total Ig conjugate. Analyses of paired serum samples from 20 seropositive dogs revealed temporal differences in titers; changes occurred during brief (1 month) or extended (greater than 4 years) sampling periods. Clinical records for 52 seropositive dogs indicated a history of intermittent lameness in 19 of these. Limb/joint disorders typically developed in dogs without IgM antibodies, suggesting manifestation during later phases of illness. A microscopic-agglutination test was used to assess cross reactivity between B burgdorferi and 20 serovars of Leptospira interrogans and biflexa. Analyses of 63 dog serum specimens with antibodies to B burgdorferi and a series of reference rabbit sera revealed minor antigenic relatedness. There was geographic clustering of dogs with antibodies to B burgdorferi in areas of south-central and southeastern Connecticut, where human Lyme disease also occurs. 相似文献
9.
L L Werner R E Halliwell R F Jackson T C Needham M Limpach 《Veterinary immunology and immunopathology》1984,7(3-4):285-292
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. 相似文献
10.
Sriwichai C Jiraudommongkol M Wutti-In Y Vongchan P 《The Journal of small animal practice》2011,52(9):476-483
Objectives : Canine allo‐ or autoantibodies are clinically important, but attachment of these immunoglobulin G (IgG) antibodies does not produce observable haemagglutination. Antibody to canine globulins is required to demonstrate sensitisation of red blood cells. Commercial reagents are available, but these often differ in sensitivity and specificity. Rabbit anticanine globulins (polyspecific) were produced for use in canine blood compatibility testing and in the investigation of immune‐mediated haemolytic anaemia. Methods : Canine sera was pooled, IgG was purified and subsequently used to immunise rabbits. A rising titre of anticanine IgG was demonstrated by indirect enzyme‐linked immunosorbent assay. Rabbit anticanine complement was isolated and investigated by agglutination of complement‐coated canine red blood cells. Both antibodies were purified and checked for crossreactivity before being combined to polyspecific anticanine globulins. The obtained reagent was used to indicate sensitised canine red blood cells and free antibodies in three different types of clinical samples, including blood for compatibility testing and that for investigation of immune‐mediated haemolytic anaemia and screening for post‐transfusion alloantibodies and was also compared to commercial Coombs’ reagent. Results : The product provided results in accordance with those from commercial Coombs’ reagent. The sensitivity for canine crossmatching was 100% and specificity for diagnosing immune‐mediated haemolytic anaemia was 87%. Clinical Significance : This product is helpful for canine crossmatching purposes and in the investigation of immune‐mediated haemolytic anaemia. 相似文献
11.
The clinical, haematological and immunological findings in 24 dogs with Coombs' positive haemolytic anaemia are described; 33% were Old English Sheepdogs. Dogs with intravascular haemolysis had a shorter history of illnesses, more severe clinical signs including vomiting, jaundice and fever, and had a poor survival rate compared to dogs with extravascular haemolysis. The anaemia was severe and regenerative in 18 dogs, and was characterised by spherocytosis and microscopic red cell agglutination, with leukocytosis. Serum IgG levels were elevated in 20 dogs, and changes in IgM, IgA, C3 and C4 were found. Antinuclear antibody was also demonstrated in 13 dogs, of which 7 were Old English Sheepdogs. It is suggested that a distinct multisystem autoimmune syndrome exists within the local Old English Sheepdog population. 相似文献
12.
Pitfalls in immunofluorescence testing in canine dermatology 总被引：1，自引：0，他引：1
Indirect immunofluorescence testing for pemphigus-like antibodies was performed on 100 dogs: 50 dogs with various nonpemphigus dermatologic diseases, 25 dogs with various nondermatologic diseases, and 25 normal dogs. One dog (generalized demodicosis) was positive for pemphigus-like antibodies at a titer of 1:10. It was concluded that canine pemphigus-like antibodies are a potential source of misinterpretation and misdiagnosis in indirect immunofluorescence testing. Direct immunofluorescence testing for IgG, IgA, IgM, and C3 was performed on the footpads of 11 normal dogs. Granular deposition of IgM at the basement membrane zone was demonstrated in 5 of the 11 dogs. It was concluded that direct immunofluorescence testing of canine footpads using only polyvalent immunoglobulin antisera or anti-IgM antisera may lead to misinterpretation and misdiagnosis in up to 45% of all dogs tested. 相似文献
13.
Familial canine dermatomyositis: clinicopathologic, immunologic, and serologic studies 总被引：2，自引：0，他引：2
K H Haupt D J Prieur A M Hargis R L Cowell T L McDonald L L Werner J F Evermann 《American journal of veterinary research》1985,46(9):1870-1875
Laboratory studies were performed on 3 Collies with familial canine dermatomyositis, 6 progeny from a breeding of 2 of the Collies (incross litter), and 4 progeny from the breeding of an affected Collie male and a normal Labrador Retriever (outcross litter). Hematologic abnormalities were leukogram changes consistent with inflammation, and, in 2 severely affected incross dogs, anemia of inflammatory disease. Serum muscle enzyme activities were not markedly increased. High concentrations of immune complexes and proportionally increased total immunoglobulin G were present in the sera of moderately and severely affected incross dogs. The same dogs had weakly positive direct Coombs' tests, and 1 affected outcross dog had a strongly positive direct Coombs' test. Rheumatoid factor was present in a severely affected incross dog. A few dogs had polyclonal globulin increases that were attributed to inflammation. Low antibody titers to canine calicivirus were present in 3 dogs and a low titer to canine coronavirus was present in 1 dog. 相似文献
14.
Christine J Piek Erik Teske Martin W van Leeuwen Michael J Day 《Acta veterinaria Scandinavica》2012,54(1):10
Background
The aim of this study was to compare a gel-based test with the traditional direct agglutination test (DAT) for the diagnosis of immune-mediated haemolytic anaemia (IMHA).Methods
Canine (n = 247) and feline (n = 74) blood samples were submitted for DAT testing to two laboratories. A subset of canine samples was categorized as having idiopathic IMHA, secondary IMHA, or no IMHA.Results
The kappa values for agreement between the tests were in one laboratory 0.86 for canine and 0.58 for feline samples, and in the other 0.48 for canine samples. The lower agreement in the second laboratory was caused by a high number of positive canine DATs for which the gel test was negative. This group included significantly more dogs with secondary IMHA.Conclusions
The gel test might be used as a screening test for idiopathic IMHA and is less often positive in secondary IMHA than the DAT. 相似文献15.
Piek CJ 《The Veterinary quarterly》2011,31(3):129-141
Idiopathic immune-mediated haemolytic anaemia (IMHA) is one of the most common immune-mediated diseases of dogs. The aim of this article is to review current knowledge of canine IMHA, its etiology, clinical presentation, diagnosis, complications, and treatment, in an attempt to establish why its outcome is still so poor. Clinical signs of anaemia develop within 3 days and dogs present with a median haematocrit of 13%, leucocytosis, a left shift, and reticulocytosis. Coagulation test results support the presence of disseminated intravascular coagulation. About 50% of dogs die in the first 2 weeks after presentation, and analysis of risk factors suggests that mortality is associated with hypercoagulability, inflammatory response, and liver and kidney failure. A positive direct agglutination test, spherocytosis, and true autoagglutination are widely accepted tests to demonstrate anti-erythrocyte antibodies, but are not yet standardized. To date, there is no evidence to support the efficacy of immunomodulators in addition to corticosteroids in the treatment of IMHA. Despite numerous investigations, the prognosis of IMHA remains dismal. There is an urgent need to validate and standardize diagnostic tests and criteria, and clinical trials might benefit from stratifying dogs by mortality risk. Analysis of samples from well-defined cases of canine IMHA might provide insight into the aetiology and pathophysiology of IMHA. 相似文献
16.
Kucinskiene G Schuberth HJ Leibold W Pieskus J 《Veterinary journal (London, England : 1997)》2005,169(2):303-307
The detection of red blood cell (RBC)-bound immunoglobulins in case of anaemia with the direct agglutination test (DAT or Coombs test) has been reported to be of low sensitivity. We therefore tested the applicability of flow cytometry for the detection of canine IgG on RBC using two different IgG-specific secondary reagents: goat-anti-dog IgG (GalphaD-IgG) and rabbit-anti-dog IgG (RalphaD-IgG). Membrane staining RBC samples were performed at 4 degrees C. Comparisons of agglutination test at 37 degrees C and 4 degrees C showed, that binding of the secondary antibodies at 4 degrees C was more sensitive compared to agglutination at 37 degrees C and the two antisera differed considerable in their agglutination activity. Binding of GalphaD-IgG and RalphaD-IgG to RBC of healthy dogs (n=15) was low and mean fluorescence intensities were taken to calculate thresholds above which RBC of patients were judged positive. As in agglutination tests, both secondary antisera displayed considerable differences (concentration-dependent binding and histogram profiles) after flow cytometric analysis. Using flow cytometry, with GalphaD-IgG 8 of 17 agglutination-negative patients were positive and RalphaD-IgG was positive with 3 of 3 agglutination-negative RBC samples. Thus, flow cytometric analysis of proved to be a sensitive technique, detecting RBC-bound canine IgG of DAT-negative patients. The results of both techniques, however, are significantly influenced by the used IgG-specific polyclonal reagents. 相似文献
17.
OBJECTIVE: To compare canine blood-typing results determined by use of the card (CARD), gel (GEL), Michigan State University (MSU), and tube (TUBE) tests. SAMPLE POPULATION: Blood samples from 23 healthy dogs. PROCEDURES: Blood samples anticoagulated with EDTA were screened by use of each blood-typing method according to manufacturers' protocols. RESULTS: Strong RBC agglutination reactions were observed with dog erythrocyte antigen (DEA) 1.1 reagents of the CARD and GEL tests as well as MSU test (only after adding Coombs' reagent) in 9 blood samples. By use of the CARD test, RBCs from 4 additional dogs agglutinated weakly; on the basis of MSU test results, these 4 dogs were classified as DEA 1.2 positive. All blood samples agglutinated with the B antigen reagent of the TUBE test. All but 2 blood samples had strong positive reactions with the DEA 4 reagent of the MSU test. All but 3 blood samples reacted with the E antigen reagent of the TUBE test. Three blood samples agglutinated with the DEA 3 reagent of the MSU test and A antigen reagent of the TUBE test. Five blood samples had strong agglutination reactions with the DEA 5 reagent of the MSU test. CONCLUSIONS AND CLINICAL RELEVANCE: Use of the CARD test allows for rapid identification of DEA 1.1 but may produce weak reactions with blood from DEA 1.2-positive dogs. The GEL test is a reliable and rapid clinical laboratory method for identification of DEA 1.1. The MSU test requires Coombs' reagent for identification of DEA 1.1 and 1.2. 相似文献
18.
Isotype-specific antibodies in horses and dogs with immune-mediated hemolytic anemia 总被引：1，自引：1，他引：0
Wilkerson MJ Davis E Shuman W Harkin K Cox J Rush B 《Journal of veterinary internal medicine / American College of Veterinary Internal Medicine》2000,14(2):190-196
Classes of antibody bound to erythrocytes were determined using direct immunofluorescence (DIF) flow cytometry in 3 horses and 12 dogs with immune-mediated hemolytic anemia (IMHA). Background levels of antibody binding were determined in samples from 12 horses and 12 dogs that were free of clinical disease. The range of nonspecific binding of a fluorescein isothiocyanate (FITC)-conjugated goat anti-equine immunoglobulin G (IgG) was 19.9–36.7%, but was eliminated by the use of the F(ab)2 fragment of FITC-conjugated goat anti-equine IgG. Background binding by other class-specific antibodies to equine and canine erythrocytes was negligible. The DIF results were compared to the direct antiglobulin (Coombs) test in 5 horses and 20 dogs with anemia. The former assay was more sensitive in dogs with IMHA than was the Coombs' test (100% versus 58%). In contrast, the Coombs' test had better specificity than the DIF assay (100% versus 87.5%, respectively). Using clinical parameters or response to therapy as the comparison, the positive and negative predictive values for the DIF test were 92% and 100% compared to the values of the Coombs' test of 100% and 62%. The DIF assay detected low levels of cells bound with antibody (<30%) in 5 dogs that were Coombs' test-negative. For both species, performance of the DIF test was independent of the prozone effect. Five dogs with IMHA had IgG and IgM on erythrocytes, 5 had IgG, and 2 had IgM. Three horses had surface-bound IgG, including a horse with suspected penicillin-induced IMHA, a foal with neonatal isoerythrolysis, and a foal with clostridial septicemia. The DIF method was valuable in monitoring the response to therapy in the foal with neonatal isoerythrolysis. 相似文献
19.
Möhr AJ Lobetti RG van der Lugt JJ 《Journal of the South African Veterinary Association》2000,71(4):232-239
This retrospective study describes 4 cases of canine babesiosis with histologically confirmed acute pancreatitis. In addition, 16 dogs with babesiosis are reported with serum amylase (>3500 U/l) and/or lipase (>650 U/l) activity elevations of a magnitude that would support a diagnosis of probable acute pancreatitis, although extra-pancreatic sources of the enzymes could not be excluded in these cases. Median time of pancreatitis diagnosis was 2.5 days post-admission, with primarily young (median age 3 years), sexually intact dogs affected. The development of pancreatitis was unrelated to the degree of anaemia at time of admission. In addition to pancreatitis, 80% of cases suffered from other babesial complications, namely icterus (13), acute respiratory distress syndrome (6), immune-mediated haemolytic anaemia (6), renal failure (3), haemoconcentration (2) and cerebral syndrome (2). Acute respiratory distress syndrome, renal failure and cerebral syndrome were associated with a poor prognosis, with 4 of the 5 dogs included in the overall 26% mortality rate having at least 1 of these complications. Haemolytic anaemia with ischaemia-reperfusion injury to the pancreas is proposed as a possible primary pathophysiological mechanism in babesial pancreatitis. Hypotensive shock, immune-mediated haemolytic anaemia, haemoconcentration and possibly altered lipid metabolism in babesiosis may also be involved. The previously postulated pro-inflammatory cytokine milieu of complicated babesiosis may underlie the progression, if not the primary initiation, of pancreatic pathology. Acute pancreatitis may represent the previously reported 'gut' form of babesiosis. 相似文献
20.
Geert Paes Dominique Paepe Evelyne Meyer Annemarie T Kristensen Luc Duchateau Miguel Campos Sylvie Daminet 《Acta veterinaria Scandinavica》2013,55(1):74