Assay validation and biological variation of serum receptor for advanced glycation end-products

Ann Clin Biochem 2008;45:518-519
doi:10.1258/acb.2008.008043
© 2008 Association for Clinical Biochemistry

 

 

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Louise F Brown and
Callum G Fraser


Department of Biochemical Medicine, NHS Tayside, Ninewells Hospital and Medical School, Dundee DD1 9SY, UK


Corresponding author: Miss Louise F Brown. Email: l.brown3{at}nhs.net


Background: The analytical performance characteristics of an enzyme-linkedimmunosorbent assay for the receptor for advanced glycationend-products (RAGE) were evaluated. The within- and between-subjectcomponents of biological variation were also estimated.

Methods: Blood was sampled from healthy volunteers into K2-ethylenediaminetetraacetic acid (EDTA) and serum separator tubes (SST) andthe stability of RAGE in whole blood, plasma and serum examined.Performance characteristics of the assay were assessed usingquality control materials. Three samples were obtained fromeach of 21 healthy volunteers one-week apart, RAGE measuredand components of biological variation estimated.

Results: RAGE concentrations in blood specimens collected into K2-EDTA and SST were stable for at least 6 hours and, after centrifugation, both plasma and serum were stable for at least 24 hours. The RAGE assay had the following characteristics: inter-assay imprecision: coefficient of variation 7.3%, working range: 26–5000 ng/L, linearity: r = 0.9977 and detection limit: 26 ng/L. Overall within- and between-subject biological variations were 14.6% and 56.5%, the index of individuality was 0.31 and the reference change value was 49.0% at P < 0.05.

Conclusion: Samples for RAGE analyses in serum or plasma can be collectedwithout significant difficulties with the assay showing acceptableanalytical performance characteristics. Conventional population-basedreference values are of limited utility in diagnosis, indicatingthat RAGE is likely to be more useful in monitoring disease.

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