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Keywords:

  • enzyme-linked sorbent assay;
  • biotin;
  • blood

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.