Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
SHR Neuro Krebs Kardio Lipid Stoffw Microb
Farrell, CJ; Soldo, J; McWhinney, B; Bandodkar, S; Herrmann, M.
Impact of assay design on test performance: lessons learned from 25-hydroxyvitamin D.
Clin Chem Lab Med. 2014; 52(11):1579-1587
Doi: 10.1515/cclm-2014-0111
Web of Science
PubMed
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- Führende Autor*innen der Med Uni Graz
- Herrmann Markus
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- Abstract:
- Current automated immunoassays vary significantly in many aspects of their design. This study sought to establish if the theoretical advantages and disadvantages associated with different design formats of automated 25-hydroxyvitamin D (25-OHD) assays are translated into variations in assay performance in practice. 25-OHD was measured in 1236 samples using automated assays from Abbott, DiaSorin, Roche and Siemens. A subset of 362 samples had up to three liquid chromatography-tandem mass spectrometry 25-OHD analyses performed. 25-OHD₂ recovery, dilution recovery, human anti-animal antibody (HAAA) interference, 3-epi-25-OHD₃ cross-reactivity and precision of the automated assays were evaluated. The assay that combined release of 25-OHD with analyte capture in a single step showed the most accurate 25-OHD₂ recovery and the best dilution recovery. The use of vitamin D binding protein (DBP) as the capture moiety was associated with 25-OHD₂ under-recovery, a trend consistent with 3-epi-25-OHD₃ cross-reactivity and immunity to HAAA interference. Assays using animal-derived antibodies did not show 3-epi-25-OHD₃ cross-reactivity but were variably susceptible to HAAA interference. Not combining 25-OHD release and capture in one step and use of biotin-streptavidin interaction for solid phase separation were features of the assays with inferior accuracy for diluted samples. The assays that used a backfill assay format showed the best precision at high concentrations but this design did not guarantee precision at low 25-OHD concentrations. Variations in design among automated 25-OHD assays influence their performance characteristics. Consideration of the details of assay design is therefore important when selecting and validating new assays.
- Find related publications in this database (using NLM MeSH Indexing)
- 25-Hydroxyvitamin D 2 - blood
- 25-Hydroxyvitamin D 2 - immunology
- 25-Hydroxyvitamin D 2 - isolation & purification
- Antibodies - immunology
- Automation -
- Chromatography, High Pressure Liquid -
- Cross Reactions -
- Humans -
- Immunoassay - instrumentation
- Immunoassay - methods
- Protein Binding -
- Tandem Mass Spectrometry -
- Vitamin D - analogs & derivatives
- Vitamin D - blood
- Vitamin D - immunology
- Vitamin D - isolation & purification
- Vitamin D-Binding Protein - chemistry
- Vitamin D-Binding Protein - metabolism
- Find related publications in this database (Keywords)
- 25-hydroxyvitamin D
- immunoassay
- liquid chromatography-tandem mass spectrometry