Pseudo‐hypoproteinemia in a hyperbilirubinemic dog with immune‐mediated hemolytic anemia

Garner, Bridget C.; Priest, Heather; Smith, J. R.

doi:10.1111/vcp.12134

Cited by 3 publications

(4 citation statements)

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“…There are 2 common analytic methods of determining plasma or serum total protein concentrations: biuret spectrophotometric assays and refractometry. A recent case report described a pseudohypoproteinemia in markedly icteric serum when the serum was assayed with a biuret total protein assay . However, the bilirubin concentration at which the interference becomes detectable was not established.…”

Section: Introductionmentioning

confidence: 99%

“…A recent case report described a pseudohypoproteinemia in markedly icteric serum when the serum was assayed with a biuret total protein assay. 1 However, the bilirubin concentration at which the interference becomes detectable was not established. In the case report, the refractometric total protein concentration did not appear to be adversely affected by the hyperbilirubinemia.…”

Section: Introductionmentioning

confidence: 99%

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Negative interference of icteric serum on a bichromatic biuret total protein assay

Gupta

Stockham

2014

Veterinary Clinical Pathol

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Background Bilirubin is stated to be a negative interferent in some biuret assays and thus could contribute to pseudohypoproteinemia in icteric samples. Objective The purpose of the study was to evaluate the magnitude of and reason for a falsely low total protein concentration in icteric serum when the protein concentration is measured with a bichromatic spectrophotometric biuret assay. Methods Commercially available bilirubin was dissolved in 0.1 M NaOH and mixed with sera from 2 dogs to achieve various bilirubin concentrations of up to 40 mg/dL (first set of samples) and 35 mg/dL (second set of samples, for confirmation of first set of results and to explore the interference). Biuret total protein and bilirubin concentrations were determined with a chemistry analyzer (Cobas 6000 with c501 module). Line graphs were drawn to illustrate the effects of increasing bilirubin concentrations on the total protein concentrations. Specific spectrophotometric absorbance readings were examined to identify the reason for the negative interference. Results High bilirubin concentrations created a negative interference in the Cobas biuret assay. The detectable interference occurred with a spiked bilirubin concentration of 10.7 mg/dL in one set of samples, 20.8 mg/dL in a second set. The interference was due to a greater secondary‐absorbance reading at the second measuring point in the samples spiked with bilirubin, which possibly had converted to biliverdin. Conclusion Marked hyperbilirubinemia is associated with a falsely low serum total protein concentration when measured with a bichromatic spectrophotometric biuret assay. This can result in pseudohypoproteinemia and pseudohypoglobulinemia in icteric serum.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Negative interference of icteric serum on a bichromatic biuret total protein assay

Gupta

Stockham

2014

Veterinary Clinical Pathol

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“…These methods can have variable performance based on the number and type of protein species present in the sample, and minor interference can be caused by peptides and free amino acids . Additionally, icterus can cause interference with the biuret method . A survey revealed that only 2.4% of 842 participating human laboratories used refractometry‐based total protein quantification for SPE .…”

Section: Methodsmentioning

confidence: 99%

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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“…Recently, pseudohypoproteinemia was reported in a hyperbilirubinemic dog. 21 The refractometer measurement was in the reference interval, whereas the biuret assay total protein concentration was decreased. The lower biuret assay total protein concentration was at-…”

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confidence: 99%

Refractometric total protein concentrations in icteric serum from dogs

Gupta¹,

Stockham²

2014

javma

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Pseudo‐hypoproteinemia in a hyperbilirubinemic dog with immune‐mediated hemolytic anemia

Cited by 3 publications

References 11 publications

Negative interference of icteric serum on a bichromatic biuret total protein assay

Negative interference of icteric serum on a bichromatic biuret total protein assay

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

Refractometric total protein concentrations in icteric serum from dogs

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