Enzyme‐linked sorbent assay to quantify d‐biotin in blood

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Abstract
An enzyme‐linked sorbent assay (ELSA) is described to determine quantitatively d‐biotin in blood. This assay utilises the strong binding of streptavidin (as opposed to avidin which gave high background reactions). A competition reaction in a microtitration plate is set up between biotin bound to the plate with biotin in the sample and streptavidin conjugated to horseradish peroxidase using tetramethylbenzidine as a marker. This ELSA can be applied to samples with a d‐biotin concentration of down to 20 ng litre−1. There was a good correlation coefficient of 0·981 between the ELSA and the standard microbiological method for d‐biotin.