Using the hemoglobin-binding Staphylococcus aureus protein IsdH to enable plasma analysis of hemolyzed blood samples
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Kirstine Lindhardt Sæderup
Abstract
Background
Intravascular hemolysis and in vitro hemolysis are prevalent contributors to failed blood sample analysis in the routine hospital laboratory. Interferences by hemoglobin in spectrophotometric and certain enzyme activity assays is the major causative factor.
Methods
By exploiting the hemoglobin-binding properties of the iron-regulated surface determinant H (IsdH) protein from Staphylococcus aureus we have developed a new method to instantly remove hemoglobin and hemoglobin-haptoglobin complexes from plasma in vitro thereby enabling the measurement of hemoglobin-sensitive analytes in hemolyzed plasma. In the present study we used an engineered IsdH mutant form conjugated to Sepharose for the efficient removal of plasma hemoglobin in concentrations up to 15 mg/mL. The high abundance of haptoglobin, which forms a tight complex with hemoglobin in plasma, did not affect the hemoglobin removal by IsdH Sepharose.
Results
Applying the method on plasma samples that beforehand were spiked with blood hemolysate re-enabled measurement of the hemolysis sensitive parameters: alkaline phosphatase, conjugated bilirubin, iron, ferritin, γ-glutamyltransferase, total thyroxine and troponin T. IsdH Sepharose-mediated hemoglobin removal also enabled measurement of hemolysis sensitive parameters in hemolyzed samples from anonymized patients.
Conclusions
In conclusion, IsdH Sepharose is a simple cost-effective pretreatment of hemolyzed samples correcting and enabling the measurement of several important hemoglobin-sensitive parameters in a way compatible with standard procedures in routine laboratories.
Funding source: Det Frie Forskningsråd
Award Identifier / Grant number: 7017-00126B
Funding source: Lundbeckfonden
Award Identifier / Grant number: R180-2014-3649
Funding source: Novo Nordisk UK Research Foundation
Award Identifier / Grant number: NNF14OC0011537
Funding statement: Det Frie Forskningsråd, Funder Id: http://dx.doi.org/10.13039/501100004836, Grant Number: 7017-00126B. Lundbeckfonden, Funder Id: http://dx.doi.org/10.13039/501100003554, Grant Number: R180-2014-3649. Novo Nordisk UK Research Foundation, Funder Id: http://dx.doi.org/10.13039/501100000329, Grant Number: NNF14OC0011537.
Acknowledgments
We would like to thank laboratory technician Malene Bille Jørgensen and laboratory technician specialist Anette Bergstedt for their excellent technical assistance.
Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.
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.
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Supplementary Material
The online version of this article offers supplementary material (https://doi.org/10.1515/cclm-2019-0381).
©2019 Walter de Gruyter GmbH, Berlin/Boston
Articles in the same Issue
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Articles in the same Issue
- Frontmatter
- Editorial
- Identification and management of spurious hemolysis: controversies, concerns and criticisms
- Reviews
- CYP24A1 and SLC34A1 genetic defects associated with idiopathic infantile hypercalcemia: from genotype to phenotype
- Meta-analysis: compared with anti-CCP and rheumatoid factor, could anti-MCV be the next biomarker in the rheumatoid arthritis classification criteria?
- Opinion Paper
- Collection, transport and storage procedures for blood culture specimens in adult patients: recommendations from a board of Italian experts
- General Clinical Chemistry and Laboratory Medicine
- Using the hemoglobin-binding Staphylococcus aureus protein IsdH to enable plasma analysis of hemolyzed blood samples
- Handling of hemolyzed serum samples in clinical chemistry laboratories: the Nordic hemolysis project
- Clinical biomarker innovation: when is it worthwhile?
- Impact of total automation consolidating first-line laboratory tests on diagnostic blood loss
- Diagnostic and prognostic value of the D-dimer test in emergency department patients: secondary analysis of an observational study
- Screening of chemical libraries in pursuit of kallikrein-5 specific inhibitors for the treatment of inflammatory dermatoses
- Comparison of five automated urine sediment analyzers with manual microscopy for accurate identification of urine sediment
- Establishment of an international autoantibody reference standard for human anti-DFS70 antibodies: proof-of-concept study for a novel Megapool strategy by pooling individual specific sera
- Only monospecific anti-DFS70 antibodies aid in the exclusion of antinuclear antibody associated rheumatic diseases: an Italian experience
- Performance evaluation of an Indoxyl Sulfate Assay Kit “NIPRO”
- Variable and inaccurate serum IgG4 levels resulting from lack of standardization in IgG subclass assay calibration
- Hematology and Coagulation
- Influence of hypertriglyceridemia, hyperbilirubinemia and hemolysis on thrombin generation in human plasma
- Reference Values and Biological Variations
- Reference intervals for serum total vitamin B12 and holotranscobalamin concentrations and their change points with methylmalonic acid concentration to assess vitamin B12 status during early and mid-pregnancy
- Cardiovascular Diseases
- Serum concentrations of free fatty acids are associated with 3-month mortality in acute heart failure patients
- Letters to the Editor
- Management of potassium results in haemolysed plasma samples at the emergency department laboratory
- Unexpected analytical interference in isavuconazole UV determination in a child in therapy with lumacaftor/ivacaftor for cystic fibrosis
- Total laboratory automation has the potential to be the field of application of artificial intelligence: the cyber-physical system and “Automation 4.0”
- 99th percentile upper reference limit of AccuTnI+3 in a Korean reference population: a multicenter study using fresh serum
- Roche Troponin T hs-STAT meets all expert opinion analytical laboratory practice recommendations for the use of the differential diagnosis of acute coronary syndrome
- A comparison of biotin interference in routine immunoassays on the Roche Cobas 8000, Beckman Coulter DXi and Siemens Advia Centaur XPT immunoassay platforms
- The importance of the methodology and sample matrix when interpreting chromogranin A results
- The sudden death of the combined first trimester aneuploidy screening, a single centre experience in Belgium
- Reply to Luksic et al. Clin Chem Lab Med 2018;56(4):574–581
