title: Activity of the liver enzyme ornithine carbamoyltransferase (OTC) in blood: LC-MS/MS assay for non-invasive diagnosis of ornithine carbamoyltransferase deficiency.
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.
authors.affiliation: .
authors.forename: Jakub
authors.initials: J
authors.lastname: Krijt
authors.affiliation: .
authors.forename: Jitka
authors.initials: J
authors.lastname: Sokolová
authors.affiliation: .
authors.forename: Pavel
authors.initials: P
authors.lastname: Ješina
authors.affiliation: .
authors.forename: Lenka
authors.initials: L
authors.lastname: Dvořáková
authors.affiliation: .
authors.forename: Martin
authors.initials: M
authors.lastname: Řeboun
authors.affiliation: .
authors.forename: Katarína
authors.initials: K
authors.lastname: Brennerová
authors.affiliation: .
authors.forename: Martin
authors.initials: M
authors.lastname: Mistrík
authors.affiliation: .
authors.forename: Jiří
authors.initials: J
authors.lastname: Zeman
authors.affiliation: .
authors.forename: Tomáš
authors.initials: T
authors.lastname: Honzík
authors.affiliation: .
authors.forename: Viktor
authors.initials: V
authors.lastname: Kožich
mesh_terms: D002138:Calibration; D002853:Chromatography, Liquid; D041321:Chromosomes, Human, X / Q000235:genetics; D015331:Cohort Studies; D057075:Enzyme Assays / Q000379:methods*; D004795:Enzyme Stability; D005260:Female; D006579:Heterozygote; D006801:Humans; D016014:Linear Models; D008099:Liver / Q000201:enzymology*; D008297:Male; D009154:Mutation; D009954:Ornithine Carbamoyltransferase / Q000097:blood* / Q000235:genetics / Q000378:metabolism; D020163:Ornithine Carbamoyltransferase Deficiency Disease / Q000097:blood* / Q000175:diagnosis* / Q000201:enzymology; D053719:Tandem Mass Spectrometry
chemical_list: D009954:Ornithine Carbamoyltransferase
keywords: enzyme deficiency; inborn errors of metabolism; liquid chromatography-mass spectrometry (LC-MS/MS); ornithine carbamoyltransferase (OTC); urea cycle