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Clinical Chemistry and Laboratory Medicine (CCLM)

Published in Association with the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)

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Validation of commercial assays for measurements of trefoil factor family peptides in serum

1 / Ebba Nexo1

1Department of Clinical Biochemistry, Aarhus University Hospital, Aarhus, Denmark

Corresponding author: Mie H. Samson, Department of Clinical Biochemistry, Aarhus University Hospital, Nørrebrogade 44 8000 Aarhus C, Denmark Phone: +45 8949 4551, Fax: +45 8949 3060

Citation Information: Clinical Chemistry and Laboratory Medicine. Volume 49, Issue 12, Pages 2057–2060, ISSN (Online) 1437-4331, ISSN (Print) 1434-6621, DOI: 10.1515/CCLM.2011.712, September 2011

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Background: Trefoil peptides (TFF1, TFF2 and TFF3) are 7–12 kDa molecules, secreted by mucin-producing epithelial cells. Increased serum concentrations have been reported in a number of pathological conditions, which warrants the need for validated commercially available assays.

Methods: We validated commercial assays for TFF1-3 and compared results obtained with our in-house assays, using serum from blood donors.

Results: Level of detection was: ≤0.008 nmol/L. Measuring ranges were: 0.032–0.51 (TFF1), 0.038–0.76 (TFF2) and 0.019–0.15 (TFF3) nmol/L. Imprecision (CV), judged from the measurement of serum pools in two levels, was below 9% (TFF2 and TFF3) but up to 18% (mean 0.41 nmol/L) for TFF1. No cross reactivity between the TFFs (concentrations >100 nmol/L) was observed. The 95% non-parametric reference intervals were: <0.0032–0.53 (TFF1), 0.099–1.4 (TFF2) and 0.086–0.87 (TFF3) nmol/L. Comparing commercial to in-house assays (n=132), showed biases explained by differences in the calibrators (TFF1 and TFF2). A number of samples showed markedly different results.

Conclusions: The commercial assays for TFF2 and TFF3 are acceptable for use on serum samples, while the TFF1 assay revealed a poor imprecision and a too narrow measuring range. Results obtained with the commercial and the in-house assays differed, partly because of differences in the calibrators employed.

Keywords: ELISA; serum; trefoil factor family peptides

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