title: Automation and validation of a fast method for the assessment of in vivo oxidative stress levels.
abstract: BACKGROUND
  The d-ROMs test for the evaluation of serum hydroperoxides (HP) is simple, reliable, and cheap. Furthermore, it can easily be adapted to automated analyzers. Changing from the manual to an automated procedure allows the simultaneous processing of a large number of samples in a greatly reduced time, avoiding manual handling of samples and reagents and reducing variability sources.
  
  
  METHODS
  This study was performed to adjust the manual procedure to a routine automated method in the clinical laboratory. We carried out the d-ROMs test in sera from 90 subjects of both sexes (34 men and 56 women) with age ranging from 20 to 80 years (mean 51+/-14 years). All subjects were free from acute or chronic inflammatory disease, immunological disease and history or evidence of malignancy. Subjects were not on vitamin and/or antioxidant therapies.
  
  
  RESULTS
  The detection limit of the assay was 40 AU. Linearity was observed up to 475 AU. The recovery ranged between 97% and 105%. Within- and between-run imprecision was <5%. The mean HP value was 304+/-8 AU, with no significant difference between men (291+/-10 AU) and women (311+/-11 AU). A significant positive correlation was observed between age and HP in the whole population (r=0.4, p=0.0002).
  
  
  CONCLUSIONS
  The automated test for the estimation of serum hydroperoxides represents a reliable and feasible procedure for increasing efficiency and reducing costs compared to the manual method, and is particularly suitable for evaluating oxidative stress in a variety of clinical conditions.
authors.affiliation: CNR Institute of Clinical Physiology, Pisa, Italy. cristina.vassalle@ifc.cnr.it
authors.forename: Cristina
authors.initials: C
authors.lastname: Vassalle
authors.forename: Claudio
authors.initials: C
authors.lastname: Boni
authors.forename: Pietro
authors.initials: P
authors.lastname: Di Cecco
authors.forename: Rudina
authors.initials: R
authors.lastname: Ndreu
authors.forename: Gian Carlo
authors.initials: GC
authors.lastname: Zucchelli
mesh_terms: D000328:Adult; D000367:Age Factors; D000368:Aged; D000369:Aged, 80 and over; D015415:Biomarkers / Q000097:blood; D001774:Blood Chemical Analysis; D019963:Clinical Chemistry Tests / Q000191:economics / Q000295:instrumentation / Q000379:methods*; D003124:Colorimetry / Q000191:economics / Q000295:instrumentation / Q000379:methods; D050171:Dyslipidemias / Q000097:blood; D005260:Female; D006801:Humans; D006861:Hydrogen Peroxide / Q000097:blood*; D006973:Hypertension / Q000097:blood; D008297:Male; D008875:Middle Aged; D018384:Oxidative Stress; D015203:Reproducibility of Results; D012737:Sex Factors
chemical_list: D015415:Biomarkers; D006861:Hydrogen Peroxide