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Activity of the liver enzyme ornithine carbamoyltransferase (OTC) in blood: LC-MS/MS assay for non-invasive diagnosis of ornithine carbamoyltransferase deficiency

  • Jakub Krijt , Jitka Sokolová , Pavel Ješina , Lenka Dvořáková , Martin Řeboun , Katarína Brennerová , Martin Mistrík , Jiří Zeman , Tomáš Honzík and Viktor Kožich EMAIL logo
Published/Copyright: January 20, 2017
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Clinical Chemistry and Laboratory Medicine (CCLM)
From the journal Clinical Chemistry and Laboratory Medicine (CCLM) Volume 55 Issue 8

Abstract

Background:

Liver enzymes are released from hepatocytes into circulation and their activity can be measured in the blood. We examined whether the plasma activity of the liver enzyme ornithine carbamoyltransferase, determined by a novel liquid chromatography-mass spectrometry (LC-MS/MS) assay, could be utilized for the detection of OTC deficiency (OTCD), an X-linked inborn error of the urea cycle.

Methods:

The plasma ornithine carbamoyltransferase (OTC) activity was assayed in the reverse reaction using isotopically labeled citrulline-d4 as a substrate and by determination of the product, ornithine-d4, by LC-MS/MS analysis.

Results:

The plasma OTC activity in the controls was in the range of 111–658 pkat/L (n=49, median 272 pkat/L), and the activity increased linearly with serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities in patients with hepatopathy. The OTC activity was subsequently determined in 32 individuals carrying mutations in the OTC gene, and OTC/ALT and OTC/AST ratios were calculated to account for the degree of hepatopathy, which is a common finding in OTCD. The OTC/ALT ratio enabled clear differentiation of OTCD hemizygotes (n=11, range 0–69×10−6) from controls (504–3440×10−6). This ratio also enabled the detection of 11 of 12 symptomatic heterozygotes (range 38–794×10−6), while this marker did not allow for reliable differentiation of asymptomatic heterozygotes (n=9) from controls.

Conclusions:

LC-MS/MS assay of plasma OTC activity enabled the detection of all hemizygous and the majority of symptomatic heterozygous OTCD patients in the tested cohort. This study demonstrates that non-invasive assay of enzymes expressed predominantly in the liver could be used as an alternative approach for diagnosing inborn errors of metabolism.

Keywords: enzyme deficiency; inborn errors of metabolism; liquid chromatography-mass spectrometry (LC-MS/MS); ornithine carbamoyltransferase (OTC); urea cycle

Acknowledgments

The authors would like to thank Ms. Tereza Kovačiková, MSc. and Ms. Karolína Kukačková, BSc. for technical help, and Ms. Michaela Křížková, MSc. for critical reading of the manuscript.

  1. Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

  2. Research funding: The General University Hospital in Prague received support from the Ministry of Health of the Czech Republic (grant IGA NT/14159-3/2013 and program RVO-VFN 64165) and from the Operational Program Prague Competitiveness (project reg. No.CZ.2.16/3.1.00/24012). The First Faculty of Medicine received institutional support from the Charles University in Prague (research program PRVOUK-P24/LF1/3 and research program UNCE 204011).

  3. Employment or leadership: None declared.

  4. Honorarium: None declared.

  5. 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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Supplemental Material:

The online version of this article offers supplementary material (DOI: https://doi.org/10.1515/cclm-2016-0715).

Received: 2016-8-10
Accepted: 2016-11-30
Published Online: 2017-1-20
Published in Print: 2017-7-26

©2017 Walter de Gruyter GmbH, Berlin/Boston

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https://doi.org/10.1515/cclm-2016-0715
Keywords for this article
enzyme deficiency; inborn errors of metabolism; liquid chromatography-mass spectrometry (LC-MS/MS); ornithine carbamoyltransferase (OTC); urea cycle

Articles in the same Issue

  1. Frontmatter
  2. Editorial
  3. Biomarkers of acute kidney injury: a step forward
  4. Reviews
  5. Biomarkers of acute kidney injury: the pathway from discovery to clinical adoption
  6. Prognostic value of glycated hemoglobin among patients with ST-segment elevation myocardial infarction: a systematic review and meta-analysis
  7. Opinion Paper
  8. Traceability in laboratory medicine: a global driver for accurate results for patient care
  9. Point
  10. To report or not to report: a proposal on how to deal with altered test results in hemolytic samples
  11. Counterpoint
  12. Reporting altered test results in hemolyzed samples: is the cure worse than the disease?
  13. Genetics and Molecular Diagnostics
  14. Early mixed hematopoietic chimerism detection by digital droplet PCR in patients undergoing gender-mismatched hematopoietic stem cell transplantation
  15. Comparison of Abbott RealTime genotype II, GeneMatrix restriction fragment mass polymorphism and Sysmex HISCL HCV Gr assays for hepatitis C virus genotyping
  16. General Clinical Chemistry and Laboratory Medicine
  17. The relationship between vacuum and hemolysis during catheter blood collection: a retrospective analysis of six large cohorts
  18. Evaluation of the Greiner Bio-One serum separator BCA Fast Clot tube
  19. Implementation and application of moving average as continuous analytical quality control instrument demonstrated for 24 routine chemistry assays
  20. Parathormone stability in hemodialyzed patients and healthy subjects: comparison on non-centrifuged EDTA and serum samples with second- and third-generation assays
  21. Association between plasma proANP and hyperuricemia in Chinese Han women: a cross-sectional study
  22. Activity of the liver enzyme ornithine carbamoyltransferase (OTC) in blood: LC-MS/MS assay for non-invasive diagnosis of ornithine carbamoyltransferase deficiency
  23. Detecting paraprotein interference on a direct bilirubin assay by reviewing the photometric reaction data
  24. Prediction of human iron bioavailability using rapid c-ELISAs for human plasma hepcidin
  25. Reference Values and Biological Variations
  26. Determination of glucose-6-phosphate dehydrogenase cut-off values in a Tunisian population
  27. Plasma levels of endothelin-1 and renal function among young and healthy adults
  28. Cancer Diagnostics
  29. A new strategy for calculating the risk of ovarian malignancy algorithm (ROMA)
  30. Laboratory characterization of leukemic cell procoagulants
  31. Diabetes
  32. Preparation, calibration and evaluation of the First International Standard for human C-peptide
  33. Hb variants in Korea: effect on HbA1c using five routine methods
  34. Letters to the Editor
  35. Pseudohyperkalemia in capillary whole-blood samples – an occasional error or a significant problem in a pediatric hospital?
  36. Elevation of creatine kinase is linked to disease severity and predicts fatal outcomes in H7N9 infection
  37. Analytical evaluation of point-of-care procalcitonin (PCT) and clinical performances in an unselected population as compared with central lab PCT assay
  38. Evaluation of an automated commercial ELISA method for calprotectin determination in pleural fluid
  39. The unfinished story of interference in thyroid hormones with Roche immunoassays: when prewashing procedures matter
  40. Effects of apixaban on prothrombin time, activated partial thromboplastin time and anti-Xa assays: a European survey
  41. Evaluation of a chemiluminescent immunoassay for urinary aldosterone on the DiaSorin LIAISON automated platform against RIA and LC-MS/MS
  42. Complex considerations when tendering for HbA1c analysers
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