title: An automated enzymatic method on the Roche COBAS MIRA S for monitoring phenylalanine in dried blood spots of patients with phenylketonuria.
abstract: OBJECTIVE
  To describe and evaluate a phenylalanine dehydrogenase-coupled enzymatic assay for blood-spot phenylalanine (Phe) automated on the COBAS MIRA S analyzer for monitoring Phe in phenylketonuria (PKU) patients, as part of a home testing program.
  
  
  RESULTS
  This method required a four-point calibration with each run and the useful range was 9.3 to 3500 micromol/L Phe. The within-run precision (CV%) was 8.8% at a mean of 77 micromol/L Phe and 5.3% at 787 micromol/L Phe. The between-run precision was 15% and 5.6% for 104 micromol/L and 748 micromol/L Phe, respectively. Blood-spot Phe determinations by this method were compared with plasma Phe determined by the Beckman system 7300 HPLC analyzer using 152 samples collected from PKU patients and 56 samples from patients without PKU. Linear regression analysis revealed the equation y = 0.933x + 14.9. The standard error of estimate (Sy.x) was 82.9 and the correlation coefficient (r) was 0.98. A positive bias, observed for the blood-spot Phe assay with specimens containing Phe concentrations below 200 micro mol/L, was not due to carryover or tyrosine.
  
  
  CONCLUSIONS
  The results indicate that this method is acceptable for monitoring blood Phe levels in PKU patients.
authors.affiliation: University of Toronto, Ontario, Canada.
authors.forename: E W
authors.initials: EW
authors.lastname: Randell
authors.forename: D C
authors.initials: DC
authors.lastname: Lehotay
mesh_terms: D000594:Amino Acid Oxidoreductases; D001774:Blood Chemical Analysis / Q000295:instrumentation / Q000379:methods* / Q000706:statistics & numerical data; D005069:Evaluation Studies as Topic; D006801:Humans; D010649:Phenylalanine / Q000097:blood*; D010661:Phenylketonurias / Q000097:blood* / Q000178:diet therapy; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D010649:Phenylalanine; D000594:Amino Acid Oxidoreductases; C022342:phenylalanine oxidase