Evaluation of hemolysis, lipemia, and icterus interference with common clinical immunoassays
-
Amir Karin
Abstract
Objectives
Hemolysis, icterus, and lipemia (HIL) are common sources of endogenous interference in clinical laboratory testing. Defining the threshold of interference for immunoassays enables appropriate reporting of their results when they are affected by HIL.
Methods
Pools of residual patient serum samples were spiked with a known amount of interferent to create samples with varying concentrations of hemolysate, bilirubin, and Intralipid that mimicked the effects of endogenous HIL. Samples were analysed on the Alinity i analyser (Abbott Diagnostics) for more than 25 immunoassays. The average recovery relative to the non-spiked sample was calculated for each interference level and was compared to a predefined allowable bias.
Results
C-peptide, estradiol, serum folate, free T4, homocysteine, insulin, and vitamin B12 were found to be affected by hemolysis, at hemoglobin concentrations between 0.3 to 20 g/L. Immunoassays for BNP, estradiol, free T3, and homocysteine were affected by icterus at conjugated bilirubin concentrations between 50 to 1,044 μmol/L. BNP, serum folate, and homocysteine were affected by Intralipid with measured triglyceride concentrations between 0.8 to 10 mmol/L. Lastly, serological immunoassays for HIV and hepatitis A, B and C were also affected by interferences.
Conclusions
Immunoassays are impacted by varying degrees of HIL interference. Some measurands, in the presence of interference, are affected in a manner not previously indicated. The data presented herein provide an independent evaluation of HIL thresholds and will be of aid to resource-limited clinical laboratories that are unable to internally verify endogenous interferences when implementing the Alinity i analyser.
-
Research funding: None declared.
-
Author contributions: All authors have accepted responsibility for the entire content of this manuscript and approved its submission.
-
Competing interests: Authors state no conflict of interest.
-
Informed consent: Not applicable.
-
Ethical approval: Not applicable.
References
1. Knezevic, CE, Ness, MA, Tsang, PHT, Tenney, BJ, Marzinke, MA. Establishing hemolysis and lipemia acceptance thresholds for clinical chemistry tests. Clin Chim Acta 2020;510:459–65. https://doi.org/10.1016/j.cca.2020.08.004.Suche in Google Scholar PubMed
2. Agarwal, S, Vargas, G, Nordstrom, C, Tam, E, Buffone, GJ, Devaraj, S. Effect of interference from hemolysis, icterus and lipemia on routine pediatric clinical chemistry assays. Clin Chim Acta 2015;438:241–5. https://doi.org/10.1016/j.cca.2014.08.008.Suche in Google Scholar PubMed
3. Lippi, G, Cadamuro, J, von Meyer, A, Simundic, AM, European Federation of Clinical Chemistry and Laboratory Medicine (EFLM) Working Group for Preanalytical Phase (WG-PRE). Practical recommendations for managing hemolyzed samples in clinical chemistry testing. Clin Chem Lab Med 2018;56:718–27. https://doi.org/10.1515/cclm-2017-1104. 29373316.Suche in Google Scholar PubMed
4. CLSI. Hemolysis, icterus, and lipemia/turbidity indices as indicators of interference in clinical laboratory analysis; approved guideline. CLSI document C56-A. Wayne, PA: Clinical Laboratory Standard Institute; 2012.Suche in Google Scholar
5. Cadamuro, J, Lippi, G, von Meyer, A, Ibarz, M, van Dongen, E, Cornes, M, et al.. European survey on preanalytical sample handling - Part 2: Practices of European laboratories on monitoring and processing haemolytic, icteric and lipemic samples. On behalf of the European federation of clinical chemistry and laboratory medicine (EFLM) working group for the preanalytical phase (WG-PRE). Biochem Med (Zagreb) 2019;29:020705. https://doi.org/10.11613/BM.2019.020705. 31223259.Suche in Google Scholar PubMed PubMed Central
6. CLSI. Interference testing in clinical chemistry; approved guideline. CLSI document EP07-A2. Wayne, PA: Clinical and Laboratory Standards Institute; 2005.Suche in Google Scholar
7. Aarsand, AK, Fernandez-Calle, P, Webster, C, Coskun, A, Gonzales-Lao, E, Diaz-Garzon, J, et al.. The EFLM Biological Variation Database [Internet]. Available from: https://biologicalvariation.eu/ [Accessed 22 Nov 2022].Suche in Google Scholar
8. Dimech, W, Vincini, G, Karakaltsas, M. Determination of quality control limits for serological infectious disease testing using historical data. Clin Chem Lab Med 2015;53:329–36. https://doi.org/10.1515/cclm-2014-0546.Suche in Google Scholar PubMed
9. Walker, S, Dimech, W, Kiely, P, Smeh, K, Francis, B, Karakaltsas, M, et al.. An international quality control programme for PRISM chemiluminescent immunoassays in blood service and blood product laboratories. Vox Sang 2009;97:309–16. https://doi.org/10.1111/j.1423-0410.2009.01218.x.Suche in Google Scholar PubMed
10. Sciacovelli, L, Lippi, G, Sumarac, Z, Del Pino Castro, IG, Ivanov, A, De Guire, V, et al., Working Group “Laboratory Errors and Patient Safety” of International Federation of Clinical Chemistry and Laboratory Medicine (IFCC). Pre-analytical quality indicators in laboratory medicine: Performance of laboratories participating in the IFCC working group “Laboratory Errors and Patient Safety” project. Clin Chim Acta 2019;497:35–40. https://doi.org/10.1016/j.cca.2019.07.007. 31295446.Suche in Google Scholar PubMed
11. Barbato, L, Campelo, MD, Pigozzo, S, Realdon, N, Gandini, A, Barbazza, R, et al.. Rejection of hemolyzed samples can jeopardize patient safety. EJIFCC 2020;31:15–20. 32256285.Suche in Google Scholar
12. Zornitzki, T, Blaychfeld-Magnazi, M, Knobler, H, Ulman, M. The effect of phlebotomy-induced hemolysis on insulin level determination. Endocr Pract 2015;21:1093–7. https://doi.org/10.4158/ep15698.or.Suche in Google Scholar
13. Garinet, S, Fellahi, S, Marlin, G, Capeau, J, Lefèvre, G, Bastard, JP. Differential interferences of hemoglobin and hemolysis on insulin assay with the Abbott Architect-Ci8200 immunoassay. Clin Biochem 2014;47:445–7. https://doi.org/10.1016/j.clinbiochem.2014.01.026. 24486652.Suche in Google Scholar PubMed
14. Hasanato, R, Brearton, S, Alshebani, M, Bailey, L, Aldugashim, S, Alothaim, A, et al.. Effects of serum indices interference on hormonal results from the Abbott Architect i2000 immunoassay analyser. Br J Biomed Sci 2015;72:151–5. https://doi.org/10.1080/09674845.2015.11665744.Suche in Google Scholar PubMed
15. La’ulu, SL, Rawlins, ML, Pfeiffer, CM, Zhang, M, Roberts, WL. Performance characteristics of six homocysteine assays. Am J Clin Pathol 2008;130:969–75. https://doi.org/10.1309/ajcp64bjipnpsqdj.Suche in Google Scholar PubMed
16. Armbruster, DA, Jirinzu, DC, Williams, JV. Evaluation of enhanced luminescence immunoenzymometric assays (LIA) for ferritin and free T4. J Clin Lab Anal 1989;3:78–83. https://doi.org/10.1002/jcla.1860030203.Suche in Google Scholar PubMed
17. Saenger, AK, Jaffe, AS, Body, R, Collinson, PO, Kavsak, PA, Lam, CSP, et al.. Cardiac troponin and natriuretic peptide analytical interferences from hemolysis and biotin: educational aids from the IFCC Committee on Cardiac Biomarkers (IFCC C-CB). Clin Chem Lab Med 2019;57:633–40. https://doi.org/10.1515/cclm-2018-0905. 30291783.Suche in Google Scholar PubMed
18. Harley, K, Bissonnette, S, Inzitari, R, Schulz, K, Apple, FS, Kavsak, PA, et al.. Independent and combined effects of biotin and hemolysis on high-sensitivity cardiac troponin assays. Clin Chem Lab Med 2021;59:1431–43. https://doi.org/10.1515/cclm-2021-0124. 33761581.Suche in Google Scholar PubMed
19. Rotenberg, M, Rubin, M, Bor, A, Meyuhas, D, Talmon, Y, Lichtenberg, D. Physico-chemical characterization of IntralipidTM emulsions. Biochim Biophys Acta Lipids Lipid Metabol 1991;1086:265–72. https://doi.org/10.1016/0005-2760(91)90169-i.Suche in Google Scholar PubMed
20. Mainali, S, Davis, SR, Krasowski, MD. Frequency and causes of lipemia interference of clinical chemistry laboratory tests. Pract Lab Med 2017;8:1–9. https://doi.org/10.1016/j.plabm.2017.02.001.Suche in Google Scholar PubMed PubMed Central
Supplementary Material
The online version of this article offers supplementary material (https://doi.org/10.1515/cclm-2022-0924).
© 2022 Walter de Gruyter GmbH, Berlin/Boston
Artikel in diesem Heft
- Frontmatter
- Editorial
- Improving access to diagnostic testing in conflict-affected areas: what is needed?
- Review
- Deciphering the role of monocyte and monocyte distribution width (MDW) in COVID-19: an updated systematic review and meta-analysis
- Opinion Paper
- From research cohorts to the patient – a role for “omics” in diagnostics and laboratory medicine?
- EFLM Paper
- The European Register of Specialists in Clinical Chemistry and Laboratory Medicine: code of conduct, version 3 – 2023
- Guidelines and Recommendations
- Cardiac troponin measurement at the point of care: educational recommendations on analytical and clinical aspects by the IFCC Committee on Clinical Applications of Cardiac Bio-Markers (IFCC C-CB)
- Genetics and Molecular Diagnostics
- A new and improved method of library preparation for non-invasive prenatal testing: plasma to library express technology
- Multiplex proteomics using proximity extension assay for the identification of protein biomarkers predictive of acute graft-vs.-host disease in allogeneic hematopoietic cell transplantation
- General Clinical Chemistry and Laboratory Medicine
- Assessment of laboratory capacity in conflict-affected low-resource settings using two World Health Organization laboratory assessment tools
- Challenge in hyponatremic patients – the potential of a laboratory-based decision support system for hyponatremia to improve patient’s safety
- Evaluation of hemolysis, lipemia, and icterus interference with common clinical immunoassays
- Serum bicarbonate stability study at room temperature – influence of time to centrifugation and air exposure on bicarbonate measurement reported according to the CRESS checklist
- Effects of lipemia on capillary serum protein electrophoresis in native ultra-lipemic material and intravenous lipid emulsion added sera
- C-reactive protein interacts with amphotericin B liposomes and its potential clinical consequences
- Evaluation of analytical performance of homocysteine LC-MS/MS assay and design of internal quality control strategy
- A reliable and high throughput HPLC–HRMS method for the rapid screening of β-thalassemia and hemoglobinopathy in dried blood spots
- Dynamics of the vitamin D C3-epimer levels in preterm infants
- Comparison of ANA testing by indirect immunofluorescence or solid-phase assays in a low pre-test probability population for systemic autoimmune disease: the Camargo Cohort
- Reference Values and Biological Variations
- LMS-based continuous reference percentiles for 14 laboratory parameters in the CALIPER cohort of healthy children and adolescents
- Indirectly determined reference intervals for automated white blood cell differentials of pediatric patients in Berlin and Brandenburg
- Infectious Diseases
- Evaluation of a high-sensitivity SARS-CoV-2 antigen test on the fully automated light-initiated chemiluminescent immunoassay platform
- Letters to the Editor
- Non-esterified fatty acids (NEFA): sample stability and effect of haemolysis and icterus
- Frozen serum sample pool should not be used as internal quality assessment for lipemia (L) index
- Stability of SARS-CoV-2 respiratory samples in non-freezing condition: importance for tropical countries under heavy diagnostic demand
- A rare case of both macro-TSH and macro-LH: laboratory analysis of the pathogenesis
- A new method for early cancer detection based on platelet transcriptomics will have low positive predictive value
- Comparison of near-infrared and UV–vis-based non-contact hematocrit prediction of dried blood spots from patients on immunosuppressants
- The diagnostics of heparin-induced thrombocytopenia in Italy and the possible impact of vaccine-induced immune thrombotic thrombocytopenia on it
- Pitfall in the analysis of the alcohol biomarkers ethyl glucuronide and ethyl sulfate by laboratory-caused contamination with disinfectants
- Growing your own food is like printing money, don’t let it make you mad as a hatter
- Congress Abstracts
- 43rd annual conference of the association of clinical biochemists in Ireland (ACBI 2021)
- 44th Annual Conference of the Association of Clinical Biochemists in Ireland (ACBI 2022)
Artikel in diesem Heft
- Frontmatter
- Editorial
- Improving access to diagnostic testing in conflict-affected areas: what is needed?
- Review
- Deciphering the role of monocyte and monocyte distribution width (MDW) in COVID-19: an updated systematic review and meta-analysis
- Opinion Paper
- From research cohorts to the patient – a role for “omics” in diagnostics and laboratory medicine?
- EFLM Paper
- The European Register of Specialists in Clinical Chemistry and Laboratory Medicine: code of conduct, version 3 – 2023
- Guidelines and Recommendations
- Cardiac troponin measurement at the point of care: educational recommendations on analytical and clinical aspects by the IFCC Committee on Clinical Applications of Cardiac Bio-Markers (IFCC C-CB)
- Genetics and Molecular Diagnostics
- A new and improved method of library preparation for non-invasive prenatal testing: plasma to library express technology
- Multiplex proteomics using proximity extension assay for the identification of protein biomarkers predictive of acute graft-vs.-host disease in allogeneic hematopoietic cell transplantation
- General Clinical Chemistry and Laboratory Medicine
- Assessment of laboratory capacity in conflict-affected low-resource settings using two World Health Organization laboratory assessment tools
- Challenge in hyponatremic patients – the potential of a laboratory-based decision support system for hyponatremia to improve patient’s safety
- Evaluation of hemolysis, lipemia, and icterus interference with common clinical immunoassays
- Serum bicarbonate stability study at room temperature – influence of time to centrifugation and air exposure on bicarbonate measurement reported according to the CRESS checklist
- Effects of lipemia on capillary serum protein electrophoresis in native ultra-lipemic material and intravenous lipid emulsion added sera
- C-reactive protein interacts with amphotericin B liposomes and its potential clinical consequences
- Evaluation of analytical performance of homocysteine LC-MS/MS assay and design of internal quality control strategy
- A reliable and high throughput HPLC–HRMS method for the rapid screening of β-thalassemia and hemoglobinopathy in dried blood spots
- Dynamics of the vitamin D C3-epimer levels in preterm infants
- Comparison of ANA testing by indirect immunofluorescence or solid-phase assays in a low pre-test probability population for systemic autoimmune disease: the Camargo Cohort
- Reference Values and Biological Variations
- LMS-based continuous reference percentiles for 14 laboratory parameters in the CALIPER cohort of healthy children and adolescents
- Indirectly determined reference intervals for automated white blood cell differentials of pediatric patients in Berlin and Brandenburg
- Infectious Diseases
- Evaluation of a high-sensitivity SARS-CoV-2 antigen test on the fully automated light-initiated chemiluminescent immunoassay platform
- Letters to the Editor
- Non-esterified fatty acids (NEFA): sample stability and effect of haemolysis and icterus
- Frozen serum sample pool should not be used as internal quality assessment for lipemia (L) index
- Stability of SARS-CoV-2 respiratory samples in non-freezing condition: importance for tropical countries under heavy diagnostic demand
- A rare case of both macro-TSH and macro-LH: laboratory analysis of the pathogenesis
- A new method for early cancer detection based on platelet transcriptomics will have low positive predictive value
- Comparison of near-infrared and UV–vis-based non-contact hematocrit prediction of dried blood spots from patients on immunosuppressants
- The diagnostics of heparin-induced thrombocytopenia in Italy and the possible impact of vaccine-induced immune thrombotic thrombocytopenia on it
- Pitfall in the analysis of the alcohol biomarkers ethyl glucuronide and ethyl sulfate by laboratory-caused contamination with disinfectants
- Growing your own food is like printing money, don’t let it make you mad as a hatter
- Congress Abstracts
- 43rd annual conference of the association of clinical biochemists in Ireland (ACBI 2021)
- 44th Annual Conference of the Association of Clinical Biochemists in Ireland (ACBI 2022)
