Parathormone stability in hemodialyzed patients and healthy subjects: comparison on non-centrifuged EDTA and serum samples with second- and third-generation assays
-
Marie-Louise Schleck
und
Etienne Cavalier
Abstract
Background:
Parathyroid hormone (PTH) stability is important. Many studies have shown divergent results between EDTA and serum, which are mainly linked to differences in protocols or cut-offs used to determine whether or not PTH remained stable. No studies have yet compared PTH stability as measured by second- and third-generation assays on the same samples in hemodialyzed patients and healthy subjects.
Methods:
Five pairs of samples (EDTA and gel tubes) were obtained in 10 hemodialyzed patients before a dialysis session and in 10 healthy subjects. One pair was centrifuged and run directly to define the “T0”. Two pairs were kept at +4°C and two pairs were kept at +25°C. They were centrifuged after 4 and 18 h. Supernatant was kept at –80°C for 1 week. All samples were measured in a single batch, on Roche Cobas and DiaSorin XL second- and third-generation PTH assays. We used three different approaches to evaluate PTH stability: Wilcoxon test, an Acceptable Change Limit (ACL) according to ISO Guide 5725-6 and a Total Change Limit (TCL) derived from the sum of biological and technical variability according to WHO.
Results:
PTH decreased in all samples. Stability of PTH was mainly dependent on the way it was evaluated. Percentages of decrease were systematically lower in EDTA vs. serum. Wilcoxon and ACL showed that PTH was no more stable after 4 h at +4°C in EDTA or serum gel tubes. None of the subjects presented a PTH decrease higher than the TCL with EDTA plasma. In serum gel tubes, PTH was unstable only when kept at 25°C for 18 h.
Conclusions:
PTH seems more stable in EDTA than in serum gel tubes but only when samples have to stay unprocessed for a long period (18 h) at room temperature (25°C), which can happen when samples are delivered from external care centers. For all the other conditions, using serum gel tubes is recommended since calcium measurement, which is necessary for a good PTH results interpretation, can be achieved on the same tube.
Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.
Research funding: None declared.
Employment or leadership: None declared.
Honorarium: None declared.
Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.
References
1. Delanaye P, Souberbielle J-C, Lafage-Proust MH, Jean G, Cavalier E. Can we use circulating biomarkers to monitor bone turnover in CKD haemodialysis patients? Hypotheses and facts. Nephrol Dial Transplant 2014;29:997–1004.10.1093/ndt/gft275Suche in Google Scholar
2. Hanon EA, Sturgeon CM, Lamb EJ. Sampling and storage conditions influencing the measurement of parathyroid hormone in blood samples: a systematic review. Clin Chem Lab Med 2013;51:1925–41.10.1515/cclm-2013-0315Suche in Google Scholar
3. Cavalier E, Plebani M, Delanaye P, Souberbielle J-C. Considerations in parathyroid hormone testing. Clin Chem Lab Med 2015;53:1913–9. Available at: www.degruyter.com/view/j/cclm.ahead-of-print/cclm-2015-0314/cclm-2015-0314.xml. Accessed: 2 Jun 2015.10.1515/cclm-2015-0314Suche in Google Scholar
4. Oddoze C, Lombard E, Portugal H. Stability study of 81 analytes in human whole blood, in serum and in plasma. Clin Biochem 2012;45:464–9. Available at: http://dx.doi.org/10.1016/j.clinbiochem.2012.01.012. Accessed: 18 Jan 2012.10.1016/j.clinbiochem.2012.01.012Suche in Google Scholar
5. Scharnhornst V, Valkenburg J, Vosters C, Vader H. Influence of preanalytical factors on the immulite intact parathyroid hormone assay. Clin Chem 2004;50:974–5.10.1373/clinchem.2003.027912Suche in Google Scholar
6. Glendenning P, Laffer LL, Weber HK, Musk AA, Vasikaran SD. Parathyroid hormone is more stable in EDTA plasma than in serum. Clin Chem 2002;48:766–7.10.1093/clinchem/48.5.766Suche in Google Scholar
7. Jane Ellis M, Livesey JH, Evans MJ. Hormone stability in human whole blood. Clin Biochem 2003;36:109–12.10.1016/S0009-9120(02)00440-XSuche in Google Scholar
8. Evans MJ, Livesey JH, Ellis MJ, Yandle TG. Effect of anticoagulants and storage temperatures on stability of plasma and serum hormones. Clin Biochem 2001;34:107–12.10.1016/S0009-9120(01)00196-5Suche in Google Scholar
9. Zwart SR, Wolf M, Rogers A, Rodgers S, Gillman PL, Hitchcox K, et al. Stability of analytes related to clinical chemistry and bone metabolism in blood specimens after delayed processing. Clin Biochem 2009;42:907–10. Available at: http://dx.doi.org/10.1016/j.clinbiochem.2009.02.010. Accessed: 26 Feb 2009.10.1016/j.clinbiochem.2009.02.010Suche in Google Scholar PubMed
10. Gutierrez O, Isakova T, Rhee E, Shah A, Holmes J, Collerone G, et al. Fibroblast growth factor-23 mitigates hyperphosphatemia but accentuates calcitriol deficiency in chronic kidney disease. J Am Soc Nephrol 2005;16:2205–15.10.1681/ASN.2005010052Suche in Google Scholar PubMed
11. Holmes DT, Levin A, Forer B, Rosenberg F. Preanalytical influences on DPC IMMULITE 2000 intact PTH assays of plasma and serum from dialysis patients. Clin Chem 2005;51:913–5.10.1373/clinchem.2004.046821Suche in Google Scholar PubMed
12. Morales García AI, Górriz Teruel JL, Plancha Mansanet MC, Escudero Quesada V, Pallardó Mateu LM. Analysis of variability in determining intact parathyroid hormone (iPTH) according to the method used to process the sample. Nefrologia 2009;29:331–5.Suche in Google Scholar
13. Parent X, Alenabi F, Brignon P, Souberbielle J-C. Delayed measurement of PTH in patients with CKD: storage of the primary tube in the dialysis unit, which temperature? Which kind of tube? Nephrol Ther 2009;5:34–40.10.1016/j.nephro.2008.04.006Suche in Google Scholar PubMed
14. Joly D, Drueke TB, Alberti C, Houillier P, Lawson-Body E, Martin KJ, et al. Variation in serum and plasma PTH levels in second-generation assays in hemodialysis patients: a cross-sectional study. Am J Kidney Dis. 2008;51:987–95.10.1053/j.ajkd.2008.01.017Suche in Google Scholar PubMed
15. Parent X, Alenabi F, Etienne E, Brignon P, Chantrel F, Meynaud-Kraemer L. Variabilité pré-analytique du dosage de la parathormone chez le patient dialysé; application à l’automate Elecsys 2010 (Roche). Ann Biol Clin (Paris) 2008;66:53–8.Suche in Google Scholar
16. Ratcliffe W, Heath D, Ryan M, Jones SR. Performance and diagnostic application of a two-site immunoradiometric assay for parathyrin in serum. Clin Chem 1989;35:1957–61.10.1093/clinchem/35.9.1957Suche in Google Scholar
17. Stokes FJ, Ivanov P, Bailey LM, Fraser WD. The effects of sampling procedures and storage conditions on short-term stability of blood-based biochemical markers of bone metabolism. Clin Chem 2011;57:138–40.10.1373/clinchem.2010.157289Suche in Google Scholar PubMed
18. Cavalier E, Delanaye P, Hubert P, Krzesinski JM, Chapelle JP, Rozet E. Estimation of the stability of parathyroid hormone when stored at −80°C for a long period. Clin J Am Soc Nephrol 2009;4:1988–92.10.2215/CJN.03970609Suche in Google Scholar PubMed PubMed Central
19. Teal TK, Wood JL, Stevens PE, Lamb EJ. Stability of bio-intact (1-84) parathyroid hormone ex vivo in serum and EDTA plasma from hemodialysis patients. Clin Chem 2004;50:1713–4.10.1373/clinchem.2004.037994Suche in Google Scholar PubMed
20. ISO Guide 30:2015. Terms and conditions used in connection with reference materials.Suche in Google Scholar
21. Cavalier E, Carlisi A, Bekaert A-C, Rousselle O, Chapelle JP, Delanaye P. New insights on the stability of the parathyroid hormone as assayed by an automated 3rd generation PTH assay. Clin Chim Acta 2012;413:353–4. Available at: http://dx.doi.org/10.1016/j.cca.2011.09.034. Accessed: 3 Oct 2011.10.1016/j.cca.2011.09.034Suche in Google Scholar PubMed
22. World Health Organization. Use of anticoagulants in diagnostic laboratory investigations. 2002.Suche in Google Scholar
23. ISO Guide 5725-6:1994. Accuracy (trueness and precision) of measurement methods and results–part 6: use in practice of accuracy values.Suche in Google Scholar
24. Gardham C, Stevens PE, Delaney MP, LeRoux M, Coleman A, Lamb EJ. Variability of parathyroid hormone and other markers of bone mineral metabolism in patients receiving hemodialysis. Clin J Am Soc Nephrol 2010;5:1261–7.10.2215/CJN.09471209Suche in Google Scholar PubMed PubMed Central
25. Fraser CG, Harris EK. Generation and application of data on biological variation in clinical chemistry. Crit Rev Clin Lab Sci 1989;27:409–37.10.3109/10408368909106595Suche in Google Scholar PubMed
26. Forsberg G, Brobjer M, Holmgren E, Bergdahl K, Persson P, Gautvik KM, et al. Thrombin and H64A subtilisin cleavage of fusion proteins for preparation of human recombinant parathyroid hormone. J Protein Chem 1991;10:517–26.10.1007/BF01025480Suche in Google Scholar PubMed
27. La’ulu SL, Straseski JA, Schmidt RL, Genzen JR. Thrombin-mediated degradation of parathyroid hormone in serum tubes. Clin Chim Acta 2014;437:191–6. Available at: http://linkinghub.elsevier.com/retrieve/pii/S000989811400326X. Accessed: 30 Jul 2014.10.1016/j.cca.2014.07.030Suche in Google Scholar PubMed
28. Cavalier E, Delanaye P, Moranne O. Variability of new bone mineral metabolism markers in patients treated with maintenance hemodialysis: implications for clinical decision making. Am J Kidney Dis 2013;61:847–8. Available at: http://www.ncbi.nlm.nih.gov/pubmed/23357107. Accessed: 26 Jan 2013.10.1053/j.ajkd.2012.12.013Suche in Google Scholar PubMed
©2017 Walter de Gruyter GmbH, Berlin/Boston
Artikel in diesem Heft
- Frontmatter
- Editorial
- Biomarkers of acute kidney injury: a step forward
- Reviews
- Biomarkers of acute kidney injury: the pathway from discovery to clinical adoption
- Prognostic value of glycated hemoglobin among patients with ST-segment elevation myocardial infarction: a systematic review and meta-analysis
- Opinion Paper
- Traceability in laboratory medicine: a global driver for accurate results for patient care
- Point
- To report or not to report: a proposal on how to deal with altered test results in hemolytic samples
- Counterpoint
- Reporting altered test results in hemolyzed samples: is the cure worse than the disease?
- Genetics and Molecular Diagnostics
- Early mixed hematopoietic chimerism detection by digital droplet PCR in patients undergoing gender-mismatched hematopoietic stem cell transplantation
- Comparison of Abbott RealTime genotype II, GeneMatrix restriction fragment mass polymorphism and Sysmex HISCL HCV Gr assays for hepatitis C virus genotyping
- General Clinical Chemistry and Laboratory Medicine
- The relationship between vacuum and hemolysis during catheter blood collection: a retrospective analysis of six large cohorts
- Evaluation of the Greiner Bio-One serum separator BCA Fast Clot tube
- Implementation and application of moving average as continuous analytical quality control instrument demonstrated for 24 routine chemistry assays
- Parathormone stability in hemodialyzed patients and healthy subjects: comparison on non-centrifuged EDTA and serum samples with second- and third-generation assays
- Association between plasma proANP and hyperuricemia in Chinese Han women: a cross-sectional study
- Activity of the liver enzyme ornithine carbamoyltransferase (OTC) in blood: LC-MS/MS assay for non-invasive diagnosis of ornithine carbamoyltransferase deficiency
- Detecting paraprotein interference on a direct bilirubin assay by reviewing the photometric reaction data
- Prediction of human iron bioavailability using rapid c-ELISAs for human plasma hepcidin
- Reference Values and Biological Variations
- Determination of glucose-6-phosphate dehydrogenase cut-off values in a Tunisian population
- Plasma levels of endothelin-1 and renal function among young and healthy adults
- Cancer Diagnostics
- A new strategy for calculating the risk of ovarian malignancy algorithm (ROMA)
- Laboratory characterization of leukemic cell procoagulants
- Diabetes
- Preparation, calibration and evaluation of the First International Standard for human C-peptide
- Hb variants in Korea: effect on HbA1c using five routine methods
- Letters to the Editor
- Pseudohyperkalemia in capillary whole-blood samples – an occasional error or a significant problem in a pediatric hospital?
- Elevation of creatine kinase is linked to disease severity and predicts fatal outcomes in H7N9 infection
- Analytical evaluation of point-of-care procalcitonin (PCT) and clinical performances in an unselected population as compared with central lab PCT assay
- Evaluation of an automated commercial ELISA method for calprotectin determination in pleural fluid
- The unfinished story of interference in thyroid hormones with Roche immunoassays: when prewashing procedures matter
- Effects of apixaban on prothrombin time, activated partial thromboplastin time and anti-Xa assays: a European survey
- Evaluation of a chemiluminescent immunoassay for urinary aldosterone on the DiaSorin LIAISON automated platform against RIA and LC-MS/MS
- Complex considerations when tendering for HbA1c analysers
Artikel in diesem Heft
- Frontmatter
- Editorial
- Biomarkers of acute kidney injury: a step forward
- Reviews
- Biomarkers of acute kidney injury: the pathway from discovery to clinical adoption
- Prognostic value of glycated hemoglobin among patients with ST-segment elevation myocardial infarction: a systematic review and meta-analysis
- Opinion Paper
- Traceability in laboratory medicine: a global driver for accurate results for patient care
- Point
- To report or not to report: a proposal on how to deal with altered test results in hemolytic samples
- Counterpoint
- Reporting altered test results in hemolyzed samples: is the cure worse than the disease?
- Genetics and Molecular Diagnostics
- Early mixed hematopoietic chimerism detection by digital droplet PCR in patients undergoing gender-mismatched hematopoietic stem cell transplantation
- Comparison of Abbott RealTime genotype II, GeneMatrix restriction fragment mass polymorphism and Sysmex HISCL HCV Gr assays for hepatitis C virus genotyping
- General Clinical Chemistry and Laboratory Medicine
- The relationship between vacuum and hemolysis during catheter blood collection: a retrospective analysis of six large cohorts
- Evaluation of the Greiner Bio-One serum separator BCA Fast Clot tube
- Implementation and application of moving average as continuous analytical quality control instrument demonstrated for 24 routine chemistry assays
- Parathormone stability in hemodialyzed patients and healthy subjects: comparison on non-centrifuged EDTA and serum samples with second- and third-generation assays
- Association between plasma proANP and hyperuricemia in Chinese Han women: a cross-sectional study
- Activity of the liver enzyme ornithine carbamoyltransferase (OTC) in blood: LC-MS/MS assay for non-invasive diagnosis of ornithine carbamoyltransferase deficiency
- Detecting paraprotein interference on a direct bilirubin assay by reviewing the photometric reaction data
- Prediction of human iron bioavailability using rapid c-ELISAs for human plasma hepcidin
- Reference Values and Biological Variations
- Determination of glucose-6-phosphate dehydrogenase cut-off values in a Tunisian population
- Plasma levels of endothelin-1 and renal function among young and healthy adults
- Cancer Diagnostics
- A new strategy for calculating the risk of ovarian malignancy algorithm (ROMA)
- Laboratory characterization of leukemic cell procoagulants
- Diabetes
- Preparation, calibration and evaluation of the First International Standard for human C-peptide
- Hb variants in Korea: effect on HbA1c using five routine methods
- Letters to the Editor
- Pseudohyperkalemia in capillary whole-blood samples – an occasional error or a significant problem in a pediatric hospital?
- Elevation of creatine kinase is linked to disease severity and predicts fatal outcomes in H7N9 infection
- Analytical evaluation of point-of-care procalcitonin (PCT) and clinical performances in an unselected population as compared with central lab PCT assay
- Evaluation of an automated commercial ELISA method for calprotectin determination in pleural fluid
- The unfinished story of interference in thyroid hormones with Roche immunoassays: when prewashing procedures matter
- Effects of apixaban on prothrombin time, activated partial thromboplastin time and anti-Xa assays: a European survey
- Evaluation of a chemiluminescent immunoassay for urinary aldosterone on the DiaSorin LIAISON automated platform against RIA and LC-MS/MS
- Complex considerations when tendering for HbA1c analysers
