Validation of a competitive ELISA for the determination of serum keratan sulphate

Citation
Tb. Nikolajsen et al., Validation of a competitive ELISA for the determination of serum keratan sulphate, ANALYT CHIM, 424(2), 2000, pp. 161-167
Citations number
28
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Spectroscopy /Instrumentation/Analytical Sciences
Journal title
ANALYTICA CHIMICA ACTA
ISSN journal
0003-2670 → ACNP
Volume
424
Issue
2
Year of publication
2000
Pages
161 - 167
Database
ISI
SICI code
0003-2670(200012)424:2<161:VOACEF>2.0.ZU;2-4
Abstract
A competitive ELISA with an inhibition step for the determination of serum keratan sulphate (KS), was validated using bovine corneal KS as standard. T he method was shown to be in statistical control. The recovery was 85% (S.D . = 2.2%), hence the obtained results had to be corrected. The limit of det ection (LOD) was 1.4 mug ml(-1). Intra-assay CV was 3.0% (n = 15) for sampl es used for method evaluation and 6.0% (n = 18) for serum samples. Inter-as say CV was 7.0% for high KS level samples (C-KS = 31.2 mug ml(-1)) and 16.0 % for low KS level samples (C-KS = 5.9 mug ml(-1)) based on the S.D.(y/x) f rom the method evaluation function and 10% (high KS level) and 16% (low KS level) based on serum samples. Validation of the method was performed using the serum samples from 102 healthy individuals. There was a statistical di fference in serum KS levels between men and women, but no age related diffe rences. Tentative biological reference intervals for men and women were est ablished. It was also shown that due to the large between-subject variabili ty in KS levels, the preferable monitoring strategy is to use individuals a s their own controls where possible. (C) 2000 Elsevier Science B.V. All rig hts reserved.