Evaluation of automated assays for immunoglobulin G, M, and A measurements in dog and cat serum

Tvarijonaviciute, Asta; Martı́nez-Subiela, Silvia; Caldín, Marco; Tecles, Fernando; Cerón, José J.

doi:10.1111/vcp.12069

Cited by 12 publications

(6 citation statements)

References 33 publications

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“…Quantification of immunoglobulins in the dog has largely been restricted to the use of the radial immunodiffusion (RID), ELISA, and automated immunoturbidometric methods; based on current literature, RID appears to be the most commonly used method in veterinary medicine for quantifying the total amount of immunoglobulin . Anecdotally, we observed that the IgA RID kit significantly overestimated immunoglobulin concentrations and had poor diagnostic utility, similar to what has been documented in human medicine .…”

Section: Introductionsupporting

confidence: 56%

Validation and method comparison of the use of densitometry to quantify monoclonal proteins in canine sera

Harris

Rout

Avery

et al. 2019

Veterinary Clinical Pathol

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Background: Densitometric quantitation using serum protein electrophoresis (SPE) is used to monitor monoclonal proteins (M-proteins) in human patients but has not been validated in the dog. Serum globulin concentrations, species-specific radial immunodiffusion (RID), and ELISAs are currently used in veterinary medicine. Objective:We aimed to compare four methods that quantify M-proteins using densitometry and biuret protein (dM-protein) measurements. We also validated the best performing method and compared it with the RID and ELISA methods for measuring canine serum M-protein. Method: Serum from six normal dogs and 83 serum samples from 46 dogs with confirmed monoclonal gammopathies were used. A spike and recovery experiment with purified monoclonal IgG and IgM, inter-run and intra-run variability, linearity under dilution, and lower limit of detection were performed. Results of commercial canine RID and ELISA kits for total class-specific immunoglobulin were compared with dM-proteins. Results: The corrected perpendicular drop gating method had <20% error for IgG/γglobulin and IgM/β-globulin M-protein quantifications. Linearity (r > .99), intra-run CV (1.1%-2.3%), and inter-run CVs (2.0%-3.5%) were acceptable. Correlation between the RID and densitometry results ranged from r = .25 to r = .88, depending on the class. The RID result was greater than that of the biuret total protein in 26/63 (41%) IgA cases. A panel of IgG, IgA, and IgM RIDs failed to correctly identify an IgM paraproteinemia in 6/6 (100%) cases. Densitometry was not comparable with any other tested method. Conclusion: Densitometric quantitation is a valid technique for measuring M-proteins in the β-and γ-globulin regions. Immunotyping via RID using the tested kit does not appear to detect IgM. Densitometry is recommended for measuring M-proteins in canine patients. K E Y W O R D S electrophoresis, immunoglobulin, monoclonal gammopathy, radial immunodiffusion

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Section: Introductionsupporting

confidence: 56%

Validation and method comparison of the use of densitometry to quantify monoclonal proteins in canine sera

Harris

Rout

Avery

et al. 2019

Veterinary Clinical Pathol

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“…Group 2 and Group 3 were evaluated together as Group 2-3 whenever possible, for comparison purposes. Moreover, for a subgroup of dogs (10/12 of Group 2 and 7/9 of Group 3) IgA (immunoglobulin), IgM, and IgG fractions were quantified, respectively, at T 0– T 100 and T 0– T 250 using the method described by Tvarijonaviciute [ 16 ]. Briefly, commercial kits (Olympus Europe GmbH) were run on an automatic analyzer (Olympus AU600, Olympus Europe GmbH, Hamburg, Germany) following the manufacturer's instruction.…”

Section: Methodsmentioning

confidence: 99%

Hematological, Biochemical, and Serological Findings in Healthy Canine Blood Donors after the Administration of CaniLeish® Vaccine

Starita

Gavazza

Lubas

2016

Veterinary Medicine International

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The aim of the study was to evaluate hematological, biochemical, and serological findings in healthy canine blood donors after the administration of CaniLeish® vaccine. Twenty-seven client-owned dogs were included in the study and arranged into 3 groups according to the vaccination stage. Complete blood count (CBC) with blood smear examination, serum biochemical profile (SBP), serum protein electrophoresis (SPE), and serological tests for L. infantum were performed at different times. Additionally, in a subgroup of dogs IgA, IgM, and IgG were quantified. No statistical significance for CBC and SBP was found. In 10.7% of cases slight hyperproteinemia occurred. In SPE absolute values β-1-globulins (Group 2 and Group 2-3) and β-2-globulins (Group 3) were found modified (P < 0.05). IgG values were statistically different (P < 0.05) 6–8 months after the third immunisation (Group 2) and IgM and IgG values were statistically different after 2 months (Group 3). IFAT positive samples were 20.8% (Group 1), 15.0% (Group 2), and 52.8% (Group 3). Speed Leish K™ tests were always negative. The modifications found were probably attributed to the development of immune or inflammatory response due to the vaccine. Administration of CaniLeish vaccine in canine blood donors could be a safe practice and did not affect their health status.

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“…Similar challenges are noted with human RID and immunoturbidometric and immunonephelometric measurements . An automated human immunoturbidometric method has been validated for canine IgG, IgM, and IgA, and feline IgG and IgM . ELISA assays or refractometry have been used in the dog, cat, and horse …”

Section: Methodsmentioning

confidence: 92%

Protein characterization using electrophoresis and immunofixation; a case‐based review of dogs and cats

Moore

Avery

2019

Veterinary Clinical Pathol

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Protein electrophoresis and immunotyping can be a useful adjunct to the standard biochemical techniques for characterizing serum and urine proteins. This paper reviews currently available and commonly used methods for diagnostic protein electrophoresis, including both agarose gel and capillary zone electrophoretic techniques and total protein assessments. Immunofixation and immunosubtraction methods for identification of immunoglobulin location and class are also presented. Practical application of quality assurance and quality control strategies in compliance with American Society of Veterinary Clinical Pathology (ASVCP) best practices are discussed. Commonly encountered serum and urine electrophoretic diagnostic patterns, including electrophoretically normal, acute‐phase protein responses, polyclonal gammopathies, restricted polyclonal/oligoclonal gammopathies, paraproteinemias (monoclonal or biclonal gammopathies), and Bence‐Jones proteinurias are also reviewed using relevant case material. Cases in which immunofixation electrophoresis are particularly useful are highlighted, and methodologies to more accurately quantify serum monoclonal proteins (M‐proteins), monitoring tests commonly used in human medicine, are discussed.

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Evaluation of automated assays for immunoglobulin G, M, and A measurements in dog and cat serum

Abstract: The automated human Ig assays showed adequate precision and accuracy with serum samples from dogs and cats. Also, they were able to discriminate different concentrations of Igs in healthy and diseased animals.

Cited by 12 publications

References 33 publications

Validation and method comparison of the use of densitometry to quantify monoclonal proteins in canine sera

Validation and method comparison of the use of densitometry to quantify monoclonal proteins in canine sera

Hematological, Biochemical, and Serological Findings in Healthy Canine Blood Donors after the Administration of CaniLeish® Vaccine

Protein characterization using electrophoresis and immunofixation; a case‐based review of dogs and cats

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