Patent ID: 8802392

Claim:
A method of determining the efficacy of a sterilization process, comprising: providing a biological sterilization indicator comprising; a housing; a container containing an aqueous liquid; a source of predetermined enzyme activity that is inactivated by a sterilant; a first indicator reagent with a first absorbance spectrum; a second indicator reagent that is converted by the source of predetermined enzyme activity to a biological derivative with a second fluorescence emission spectrum; wherein the first absorbance spectrum includes detectable absorbance in at least a portion of wavelengths of the second fluorescence emission spectrum; a substrate that receives and concentrates the first indicator reagent from the aqueous liquid; providing an instrument that detects the biological derivative, wherein the instrument comprises an optical path; exposing the predetermined enzyme activity to the sterilant; bringing the predetermined enzyme activity, the first indicator reagent, the second indicator reagent, and the substrate into fluid communication with the aqueous liquid; and detecting a quantity of fluorescene from the biological derivative in the aqueous liquid; wherein detecting the quantity of fluorescence from the biological derivative comprises using the instrument; wherein detecting the quantity of fluorescence comprises positioning the housing in the optical path passes through the aqueous liquid but does not intersect the substrate; wherein the quantity of fluorescence is an indication of the efficacy of the sterilization process.