Homocysteine measurement by Vitros Microtip homocysteine assay.
BACKGROUND
Elevated circulating total homocysteine is an independent vascular risk factor. Enzymatic homocysteine measurements represent an alternative to HPLC- or immunochemistry-based assays, suitable for automation. Here, we report on analytical performance of a commercial cystathionine beta-synthase-based assay, for use on Vitros automated analyzers.


METHODS
Linear range, limit of detection and analytical sensitivity were inferred from duplicate measurements of homocystine standard solutions (1-65 micromol/L). Imprecision was assessed using commercial controls according to NCCLS EP5-A2 and accuracy using NIST-SRM1955 reference material. Agreement with a clinically validated HPLC method was examined on 207 patient samples.


RESULTS
The enzymatic assay was linear from 1 to 90 micromol/L homocysteine. Total (within-day) imprecision ranged from 4.5 (3.9)% to 2.8 (1.6)% at homocysteine 9.7-43.2 micromol/L. Accuracy was acceptable at 8.9 and 17.7 micromol/L homocysteine, with +6.4% and -1.2% bias, respectively, but showed substantial negative bias (-20.1%) at 4.0 micromol/L. High triglycerides (19.8 micromol/L) negatively interfered. The enzymatic method was slightly less sensitive than the HPLC method (limit of detection 0.7 and 0.2 micromol/L, respectively) but correlated well with the latter (r2=0.9997, slope=1.04, intercept=-0.66 micromol/L) and was more precise (p<0.05).


CONCLUSIONS
The Vitros homocysteine assay met the CLIA Desirable Analytical Quality Specifications at homocysteine > or = 9 micromol/L. Its analytical performance and suitability for automation make the Vitros assay an analytically acceptable alternative to HPLC-based methods.
Martens
Geert A
GA
Department of Clinical Chemistry and Radioimmunology, Universitair Ziekenhuis Brussel, Brussels, Belgium. geert.martens@uzbrussel.be
De Nayer
Joeri
J
De Smet
Dieter
D
Couck
Pedro
P
Gorus
Frans
F
Gerlo
Erik
E
D016477:Artifacts; D002851:Chromatography, High Pressure Liquid; D006710:Homocysteine / Q000097:blood*; D006801:Humans; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D006710:Homocysteine