Patent ID: 9476873
Filing Date: 2016-10-25
CPC Classification: G01N

Claim Text:
1. A method of measuring 25(OH) vitamin D (25OHD) in a mammalian fluid sample comprising: (a) contacting the sample comprising 25OHD with a dissociation buffer comprising a release reagent consisting of a mixture of 1-butyl-4-methylpyridinium chloride, 1-ethyl-3-methylpyridinium ethyl sulfate, methyl β-cyclodextrin and sodium salicylate in a well of a microtiter plate; (b) contacting the mixture of step (a) with (i) add-in 25OHD conjugated to an enzyme and an antibody that specifically binds to 25OHD; (c) washing the well to remove add-in 25OHD that is not specifically bound; (d) adding a substrate of said enzyme conjugated to add-in 25OHD; (e) quantitatively measuring a signal produced by the reaction of substrate and enzyme in the well; and (f) comparing the measured signal to a standard curve created with known quantities of 25OHD3, 25OHD2, or a mixture thereof, to determine the amount of 25OHD in the sample.