Lise Pedersen1, Erling Birkemose2, Charlotte Gils3, Sara Safi2, Mads Nybo2. 1. Department of Clinical Biochemistry, Holbæk Hospital, Holbæk, Denmark. 2. Department of Clinical Biochemistry and Pharmacology, Odense University Hospital, Odense, Denmark. 3. Department of Clinical Immunology and Biochemistry, Lillebaelt Hospital, Vejle, Denmark.
Abstract
BACKGROUND: Calprotectin, a complex of calcium-binding proteins, is abundant in granulocytes. Increased levels of plasma calprotectin have been found in patients with inflammatory and autoimmune diseases. However, a number of preanalytical factors may affect calprotectin measurement in blood samples. METHODS: Twelve blood samples [4 tubes, 1 each of lithium-heparin (Li-heparin), EDTA, and serum] were drawn from each of 14 healthy individuals. To evaluate the effect of temperature and storage time in the lag time between collection and centrifugation, samples were kept for 2 h at 4 °C, 20 °C, or 37 °C, before centrifugation. Leukocyte, neutrophil, and monocyte counts were measured in EDTA samples on a Sysmex XN-10 hematology analyzer to investigate the relationship between calprotectin concentrations and the granulocyte count. RESULTS: Calprotectin measurements in EDTA samples were not influenced by temperature or time lag between collection and analysis. Compared to EDTA plasma, significantly higher calprotectin concentrations were found in serum and Li-heparin plasma samples. Furthermore, calprotectin concentrations increased in serum and Li-heparin samples when stored at higher temperatures. There was a linear relationship between the serum calprotectin concentration and neutrophil count in EDTA whole blood. CONCLUSIONS: EDTA is the most suitable anticoagulant for determination of calprotectin in plasma, as this sample matrix does not seem to be affected by temperature or time between sample collection and analysis. Of particular note, neutrophil activation by either clotting or centrifugation should be avoided during the preanalytical process.
BACKGROUND: Calprotectin, a complex of calcium-binding proteins, is abundant in granulocytes. Increased levels of plasma calprotectin have been found in patients with inflammatory and autoimmune diseases. However, a number of preanalytical factors may affect calprotectin measurement in blood samples. METHODS: Twelve blood samples [4 tubes, 1 each of lithium-heparin (Li-heparin), EDTA, and serum] were drawn from each of 14 healthy individuals. To evaluate the effect of temperature and storage time in the lag time between collection and centrifugation, samples were kept for 2 h at 4 °C, 20 °C, or 37 °C, before centrifugation. Leukocyte, neutrophil, and monocyte counts were measured in EDTA samples on a Sysmex XN-10 hematology analyzer to investigate the relationship between calprotectin concentrations and the granulocyte count. RESULTS: Calprotectin measurements in EDTA samples were not influenced by temperature or time lag between collection and analysis. Compared to EDTA plasma, significantly higher calprotectin concentrations were found in serum and Li-heparin plasma samples. Furthermore, calprotectin concentrations increased in serum and Li-heparin samples when stored at higher temperatures. There was a linear relationship between the serum calprotectin concentration and neutrophil count in EDTA whole blood. CONCLUSIONS:EDTA is the most suitable anticoagulant for determination of calprotectin in plasma, as this sample matrix does not seem to be affected by temperature or time between sample collection and analysis. Of particular note, neutrophil activation by either clotting or centrifugation should be avoided during the preanalytical process.
Authors: Nancy E Cornish; Nancy L Anderson; Diego G Arambula; Matthew J Arduino; Andrew Bryan; Nancy C Burton; Bin Chen; Beverly A Dickson; Judith G Giri; Natasha K Griffith; Michael A Pentella; Reynolds M Salerno; Paramjit Sandhu; James W Snyder; Christopher A Tormey; Elizabeth A Wagar; Elizabeth G Weirich; Sheldon Campbell Journal: Clin Microbiol Rev Date: 2021-06-09 Impact factor: 50.129