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Immunology

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IMMUNOLOGY / SEROLOGY TESTS:

Test Name

 Sample Volume Canine Feline Equine Other Price
Antinuclear Antibody Antinuclear Antibody (ANA) testing by indirect fluorescent antibody (IFA) is used for detecting ANA in serum from canine, feline, and equine patients. These antibodies may be associated with systemic lupus erythematosus (SLE) and other autoimmune disorders.

Patient serum is diluted and incubated with the substrate slide containing mouse kidney cells which have prominent nuclei. After rinsing unbound serum from the slide, species specific anti-IgG labeled with FITC is added, incubated, washed and observed under a fluorescence microscope.

Results are reported as negative (<1:10) or positive with the titer and may include a description of the pattern of fluorescence such as homogeneous, peripheral, speckled, or nucleolar. 		Serum 0.5mL         $32.00
Bacterial ID by PCR w/Sequencing (Bacteria Isolate) EDTA Blood 0.5mL         $53.00
Bartonella PCR This test detects the presence of Bartonella spp.organisms in DNA extracted from blood.

The result is negative or positive with species identification based upon sequence analysis. 		EDTA Blood 0.5mL         $32.00
Bartonella henselae Titer by IFA Detects the presence of antibodies reactive with bartonella organisms.

Patient serum is diluted and incubated on a slide with B. henselae infected cells. After rinsing away unbound serum, anti-canine IgG labeled with FITC is added, incubated, and the slide washed. Canine antibody bound to bartonella is visualized using fluorescent microscopy.

The result is reported as a titer. Reactivity ≥ 1:50 is considered positive for the presence of antibody.		 Serum 0.5mL         $37.00
Blood Typing Blood Typing of canine and feline whole blood is done to identify red blood cell antigens. This lab utilizes the typing cards. The test is based upon an agglutination of antibody to type A, or type B antigen expressed on the red blood cell (RBC) surface of cats or the DEA1.1 antigen in dog red blood cells. Further testing on canine blood can be performed if a sample is determined to be negative for DEA 1.1 and involves the incubation of the washed RBCs with an antibody directed against DEA 1.X. This haemagglutination assay identifies dogs that are DEA 1.2 or DEA 1.1 positive. EDTA Blood 0.5mL     $32.00
Coombs test This test is used to detect antibody and/or complement bound to erythrocytes and is helpful for the diagnosis of immune-mediated hemolytic anemia (IMHA). The Coombs reagent contains anti- IgM, IgG, and C3 specific to the species. Red blood cells incubated with antibody are observed under an agglutination mirror and or under a microscope to determine if there is clumping or large aggregates of cells. The test is carried out at four dilutions of Coombs reagent, incubated at 37 degrees and at 4 degrees.

The result is reported as negative or positive with the Coombs titer.		 EDTA Blood 0.5mL         $21.00
Crossmatch EDTA Blood and Serum 0.5mL         $32.00
Crossmatching is performed to determine compatibility between a patient recipient and a blood cell and or plasma donor. Crossmatching is used to detect antibodies directed against red blood cells. A compatible crossmatch does not necessarily indicate that the donor and recipient have the same blood type nor does it detect platelet or granulocyte antibodies. Crossmatching should be performed prior to all transfusions especially if there is a history of prior blood exposure. Cats require crossmatching due to the dangerous consequences of transfusing the wrong blood type and the presence of naturally occurring antibodies.

The procedure requires an anticoagulated blood (EDTA purple top) tube and/ or a coagulated (red top) tube of blood from recipient and donor. A major crossmatch refers to the transfusion of donor red blood cells (RBCs) and a minor crossmatch refers to the transfusion of donor plasma. After washing the RBCs in a phosphate buffered saline solution the cells are incubated with either patient or recipient serum for thirty minutes.

The result is reported as compatible or incompatible. Ehrlichia canis realtime PCR
 EDTA Blood 0.2mL         $37.00
Ehrlichia canis Titer by IFA Test for the presence of antibodies reactive with E. canis.

Patient serum is diluted and incubated on a slide with fixed E.canis in host cells. After rinsing away unbound serum, anti-canine IgG labeled with FITC is added, incubated, and the slide washed. Canine antibody bound to E.canis organisms is visualized using fluorescent microscopy.

A positive result suggests exposure to E.canis.

The result is reported as an antibody titer.		 Serum 0.5mL         $37.00
FIV by PCR  A nested polymerase chain reaction (PCR) is used to detect the presence of the proviral form of feline immunodeficiency virus (FIV) in DNA extracted from blood. The PCR product is visualized by agarose gel electrophoresis.

Viral heterogeneity can result in false negative results do to a lack of homology between primers and their viral targets.

Results are reported as positive or negative with a cautionary note about false negative results.		 EDTA Blood 0.5mL         $32.00
Genetic Bird Sexing by PCR A polymerase chain reaction (PCR) technique, targeting the chromo-helicase DNA-binding protein (CHD) gene, is used to determine gender in birds. DNA is extracted from samples as small as 5ul of whole blood. Gender is determined following digestion of the PCR product using the restriction enzyme HaeIII or EcoRI followed by agarose gel electrophoresis.

Results are reported as male or female.		 EDTA Blood 0.2mL         $27.00
HemobartonellaHemotrophic mycoplasmas infect erythocytes in cats and can be difficult to diagnose morphologically. Real-time PCR is used to detect the organisms with a high level of sensitivity. Two different probes are used to distinguish between M. haemofelis, a pathogenic organism, and M.haemominutium, considered to be a less pathogenic organism. RT-PCR is faster than standard PCR tests which require the products be run on a gel following the reaction.

The result is reported as positive or negative.		 EDTA Blood 0.5mL         $37.00
IgG Concentration and Canine IgA Radial immunodiffusion is used to measure IgG concentrations in sera from dogs, camelids, and horses. This antigen-antibody interaction results in the formation of a zone of precipitation. The diameter of the zone is proportional to the concentration of IgG in the sample. A standard curve is generated by plotting the diameters of IgG standards. The precipitation formed by the patient sample is then measured and its concentration is extrapolated from the standard curve. The RID test is useful to determine if a failure of passive transfer (FPT) is likely to have occurred. The test, however, requires an overnight incubation.

The result is reported as an IgG concentration in mg/dL.		 Serum 0.2mL         $33.00
Immunophenotyping -- CD4, CD8, B Cells by flow cytometry, Aspirate Monoclonal antibodies are used to enumerate populations of canine and feline leukocytes. Lymphocytes are routinely examined to assist in the diagnosis of immunodeficiences and clonal expansion of malignant populations of cells as occurs in leukemias and lymphosarcomas. Samples include blood or aspirates, most often from lymph nodes or other tissue suspected of containing malignant lymphoid cells.

Results are reported as percentages of cell populations defined by monoclonal antibodies.		 EDTA Blood 0.5mL         $53.00
Lyme (B. burgdorferi)Disease Western Blot Borrelia burgdorferi antigens that have been resolved by electrophoresis and transferred to nitrocellulose membrane strip are reacted with patient serum. The presence of bound antibodies is visualzed with a secondary anti-IgG followed by an insoluble substrate.. The size of each antibody reactive antigen is compared to a template to determine band identity. Outer surface protein (OSP) A is expressed in the midgut of infected ticks. The expression of OspC occurs when ticks are in a mammalian host. Infected animals have a distinct pattern of antigen reactivity. This can be used to distinguish them from vaccinated animals and animals exposed to cross-reactive organisms.

Results are reported as positive or negative.		 Serum 0.2mL         $53.00
Mycoplasma haemofelis, Hemobartonella by Real time PCR Hemotrophic mycoplasmas infect erythocytes in cats and can be difficult to diagnose morphologically. Real-time PCR is used to detect the organisms with a high level of sensitivity. Two different probes are used to distinguish between M. haemofelis, a pathogenic organism, and M.haemominutium, considered to be a less pathogenic organism. RT-PCR is faster than standard PCR tests which require the products be run on a gel following the reaction.

The result is reported as positive or negative.		 EDTA Blood 0.5mL         $37.00
Neutrophil Function Test for ability of neutrophils to phagocytize opsonized bacteria and to generate an oxidative burst in response to a stimulus. This test requires fresh heparinized whole blood. Flow cytometry is used to enumerate the percentage of neutrophils with normal function. The test can be used for any mammalian species, however, a normal range is only available for dogs.

The result is reported as normal or abnormal and the percentage of neutrophils with normal activity.		 Heparin 1.0mL         $116.00
Protein Electrophoresis Proteins in serum, urine, or other fluids are resolved by electrophoresis and the concentration of each fraction determined by scanning densitometry. They are divided into albumin, alpha, beta , and gamma fractions. Different species of animals have different numbers of zones, alpha-1 and alpha-2, pre-albumin, etc. Monoclonal gammopathies and the ratio of albumin to gamma regions are examples of diagnostic patterns which can aid in diagnosing diseases.

Results are reported as the concentration in fraction and a densitometry scan is included.		 Serum 0.5mL         $32.00
Red Blood Cell Antibody by Flow Cyt. EDTA Blood 0.5mL         $37.00
Platelet Surface Antibody Flow Cyt. EDTA Blood 0.5mL         $37.00
Rheumatoid Factor The Canine Rheumatoid Factor (CRF) test is used to test for the presence of CRF in canine serum. Latex beads are coated with an antigen which binds to the CRF antibody present in rheumatoid arthritis.

Results are reported as negative or positive.		 Serum 0.5mL         $21.00
Tapeworm Antibody ELISA The equine tapeworm ELISA is used to detect antibodies against equine tapeworm (Anoplocephala perfoliata) antigens. The antibody concentration is compared to known positive and negative controls.

Results are reported as negative or positive and the percent reactivity compared to the positive control is included. 		Serum 0.5mL         $21.00

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University of Tennessee College of Veterinary Medicine
Department of Comparative Medicine
Clinical Immunology Service, RM A-239
2407 River Drive
Knoxville, TN 37996-4500

 


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