Reduction of dilution error in ELISAs using an internal standard
| Autor: | Thorsten Verch, Christopher Roselle, Mary Shank-Retzlaff |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Accuracy and precision Serial dilution Clinical Biochemistry Analytical chemistry Indicator Dilution Techniques Enzyme-Linked Immunosorbent Assay 01 natural sciences Fluorescence Analytical Chemistry Reduction (complexity) 03 medical and health sciences medicine General Pharmacology Toxicology and Pharmaceutics Fluorescent Dyes Chromatography medicine.diagnostic_test Chemistry 010401 analytical chemistry General Medicine Acridine Orange 0104 chemical sciences Dilution Medical Laboratory Technology 030104 developmental biology Dilution ratio Immunoassay |
| Zdroj: | Bioanalysis. 8:1451-1464 |
| ISSN: | 1757-6199 1757-6180 |
| Popis: | Background: Dilution bias is a major cause of immunoassay variability due to the lack of an internal standard to determine the true versus the expected dilution value. Methodology: We used an internal control to measure dilution bias in an ELISA. Acridine-orange was added at the first dilution step and monitored throughout dilutions. Assay results were corrected using the fluorescent signal ratio between samples and reference. Acridine dilution correlated with analyte-specific assay measurements (R2 = 0.987). Correction of assay results with the measured dilution factor improved both accuracy and precision resulting in a reduction of >50% %CV reduction. Conclusion: Dilution correction can significantly improve accuracy and precision of immunoassays. Additional control strategies may further mitigate other sources of variability. |
| Databáze: | OpenAIRE |
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