Abstract | BACKGROUND: METHODS: A two-site ELISA test was examined for measurement matrix, imprecision, limits of blank, detection, and quantitation, as well as linearity, high-dose hook effect, storage stability, cross-reactivity with nine similar compounds, interference with 22 common medications and icterus, lipemia and hemolysis, all in accordance with CLSI guidance. Also the effects of human anti-mouse antibodies and rheumatoid factor on recovery of galectin-3 were investigated. The reference interval was determined for 1092 ostensibly healthy individuals. The association of galectin-3 concentrations with an outcome of mortality was examined in a preliminary analysis of 129 acute decompensated heart failure patients. RESULTS:
Galectin-3 results were equivalent when measured in serum or EDTA plasma. Imprecision studies demonstrated that the total CV was <10% at a low concentration of 6 ng/ mL, 7% near the mid-level concentration of 21 ng/mL, and 15% at the high level of 70 ng/mL. The limit of blank was 0.86 ng/mL, the limit of detection was 1.13 ng/mL, and the limit of quantitation was 0.96 ng/mL. The linear measurement range was 0.96-130 ng/mL and there was no high-dose hook at levels up to 500 ng/mL. No cross-reactivity with nine compounds structurally related to galectin-3 and no interference from 22 common medications, icterus or lipemia was found. Hemolysis and presence of human anti-mouse antibodies or rheumatoid factor were found to be potential sources of interference. Samples can be stored for up to 15 days at either 22-28 degrees C or 2-8 degrees C before analysis; measurements are stable after storage at -20 degrees C or -70 degrees C for at least 6 months and through six freeze-thaw cycles. The 90th, 95th and 97.5th percentile of the normal reference interval was 17.6, 20.3 and 22.1 ng/mL, respectively. Galectin-3 in the acute decompensated heart failure patients ranged from 4.0 to 75 ng/mL; using a cutpoint of 22.1 ng/mL in an unadjusted analysis, galectin-3 values were associated with an outcome of death (p=0.035). Galectin-3 is associated with severity of heart failure as indicated by NYHA Class (p-value for trend, 0.017). CONCLUSION: This ELISA for galectin-3 measurement demonstrated acceptable analytical characteristics and was associated with mortality in a preliminary unadjusted analysis.
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Authors | Robert H Christenson, Show-Hong Duh, Alan H B Wu, Andrew Smith, Gyorgy Abel, Christopher R deFilippi, Shunguang Wang, Aram Adourian, Carol Adiletto, Peter Gardiner |
Journal | Clinical biochemistry
(Clin Biochem)
Vol. 43
Issue 7-8
Pg. 683-90
(May 2010)
ISSN: 1873-2933 [Electronic] United States |
PMID | 20153309
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Copyright | 2010 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved. |
Chemical References |
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Topics |
- Aged
- Animals
- Enzyme-Linked Immunosorbent Assay
(methods)
- Female
- Galectin 3
(blood)
- Heart Failure
(blood, diagnosis)
- Humans
- Limit of Detection
- Male
- Mice
- Middle Aged
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