title: Analytical performance validation of a coronary heart disease risk assessment multi-analyte proteomic test.
abstract: BACKGROUND
  Coronary heart disease (CHD) remains prevalent despite efforts to improve CHD risk assessment. The authors developed a multi-analyte immunoassay-based CHD risk assessment (CHDRA) algorithm, clinically validated in a multicenter study, to improve CHDRA in intermediate risk individuals.
  
  
  OBJECTIVE
  Clinical laboratory validation of the CHDRA biomarker assays' analytical performance.
  
  
  METHODS
  Multiplexed immunoassay panels developed for the seven CHDRA assays were evaluated with donor sera in a clinical laboratory. Specificity, sensitivity, interfering substances and reproducibility of the CHDRA assays, along with the effects of pre-analytical specimen processing, were evaluated.
  
  
  RESULTS
  Analytical measurements of the CHDRA panel proteins (CTACK, Eotaxin, Fas Ligand, HGF, IL-16, MCP-3 and sFas) exhibited acceptable accuracy (80 - 120%), cross-reactivity (< 1%), interference (< 30% at high concentrations of bilirubin, lipids, hemoglobin and HAMA), sensitivity and reproducibility (< 20% CV across multiple runs, operators and instruments). Recoveries from donor sera subjected to typical clinical laboratory pre-analytical conditions were within 80 - 120%. The pre-analytical variables did not substantively impact the CHDRA scores.
  
  
  CONCLUSIONS
  The CHDRA panel analytical validation in a clinical laboratory meets or exceeds the specifications established during the clinical utility studies. Risk score reproducibility across multiple test scenarios suggests the assays are not susceptible to clinical laboratory pre-analytical and analytical variation.
authors.affiliation: Aviir, Inc., Irvine, CA 92618, USA.
authors.forename: Niamh
authors.initials: N
authors.lastname: Nolan
authors.forename: Lilian
authors.initials: L
authors.lastname: Tee
authors.forename: Swathi
authors.initials: S
authors.lastname: Vijayakumar
authors.forename: Ivana
authors.initials: I
authors.lastname: Burazor
authors.forename: Evangelos
authors.initials: E
authors.lastname: Hytopoulos
authors.forename: William H
authors.initials: WH
authors.lastname: Biggs
authors.forename: Michael
authors.initials: M
authors.lastname: Beggs
authors.forename: Cynthia
authors.initials: C
authors.lastname: French
authors.forename: Douglas S
authors.initials: DS
authors.lastname: Harrington
mesh_terms: D015415:Biomarkers / Q000097:blood; D001798:Blood Proteins / Q000032:analysis*; D003327:Coronary Disease / Q000097:blood*; D006801:Humans; D007118:Immunoassay; D040901:Proteomics / Q000379:methods; D015203:Reproducibility of Results; D018570:Risk Assessment / Q000379:methods; D012680:Sensitivity and Specificity; D013048:Specimen Handling
chemical_list: D015415:Biomarkers; D001798:Blood Proteins