Effects of Hemolysis on Serum Chemistry Analytes in Ratites

Abstract

Since hemolysis has been a common problem in submitted ratite serum samples, a study was performed to determine interference by hemolysates. Nine chemistry analytes including glucose, total protein, albumin, creatine kinase (CK), gamma-glutamyl transferase (GGT), aspartate aminotransferase (AS7), calcium (Ca), phosphorus, and uric acid were evaluated on a wet reagent analyzer (Ciba Corning Express 550) and on a dry slide reagent analyzer (VetTest 8008). In emus, increasing hemoglobin concentrations increased total protein, albumin, and CK for both analyzers. With increasing hemoglobin concentrations, the Ciba Corning 550 analyte values were increased for AST, Ca, and uric acid and decreased for glucose and phosphorus, the opposite effect was seen in values from the VetTest 8008. GGT levels were variable or sometimes undetectable. Changes in ostrich analytes with hemolysis were similar to emus using the same analyzer. The effects of serum hemolysis often differed in magnitude and direction between the two chemistry analyzers. Interferographs were constructed to aid in rapid assessment of the effects of hemolysis on submitted serum samples.

Effect of Hemolysis on Plasma Protein Levels and Plasma Electrophoresis in Birds

“…Total protein concentration measurement may therefore be prone to both addition and optical interferences, because hemoglobin itself is a protein (Pontet, 2000), and the Biuret reaction used to carry on this measurement was based on an endpoint assay read at 553 nm. This phenomenon has been quite well documented in birds (Andreasen et al, 1996(Andreasen et al, , 1997. Hemolysis is described to induce artifactually higher total protein values (Lippi et al, 2006), even if this analysis is not extremely sensitive to interference by hemoglobin (Andreasen et al, 1996(Andreasen et al, , 1997.…”

“…This phenomenon has been quite well documented in birds (Andreasen et al, 1996(Andreasen et al, , 1997. Hemolysis is described to induce artifactually higher total protein values (Lippi et al, 2006), even if this analysis is not extremely sensitive to interference by hemoglobin (Andreasen et al, 1996(Andreasen et al, , 1997. However, effects of this interference can vary from one chemical analyzer to another (Andreasen et al, 1996(Andreasen et al, , 1997Meinkoth and Allison, 2007).…”

“…As described in other studies on avian protein electrophoresis, the A/G ratio was calculated by dividing the sum of prealbumin and albumin by the sum of the globulin fractions (Lumeij, 1987;Cray and Tatum, 1998). Effect of hemolysis in electrophoresis patterns was investigated by comparing AUCs for each fraction rather than protein concentration values, because in hemolyzed samples total protein concentrations are often erroneous (Andreasen et al, 1996(Andreasen et al, , 1997.…”

“…Hemolysis is recognized as an important interference factor in human laboratory medicine (Kroll and Elin, 1994); the release of hemoglobin and other intracellular components from erythrocytes can alter electrophoretic results. Release of hemoglobin usually gives a characteristic pink-to-red tinge to the plasma or the serum (Kirschbaumweg, 2001;Lippi et al, 2006) and is a common phenomenon in both human and veterinary medicine (Andreasen et al, 1996(Andreasen et al, , 1997Benlakehal et al, 2000;Thomas, 2001). In birds, in vivo hemolysis is relatively rare (0.3% of the samples) and can occur as a result of acute lead toxicosis or the ingestion of crude oil by shore birds (Fry and Addiego, 1987;Fudge, 2000).…”

“…Published information of the effect of hemolysis on serum chemistry for birds is limited to two studies (Andreasen et al, 1996(Andreasen et al, , 1997. In humans and birds, hemolysis can lead to artifactual high values for plasma or serum total protein concentration measurements (Andreasen et al, 1996(Andreasen et al, , 1997Pontet, 2000;Vermer et al, 2007). There is less information for the influence of hemolysis in agarose gel protein electrophoresis.…”