Gotham Biotech has developed strong collaborative associations with Marc R. Couturier, PhD at
Blastomycesdermatitidis is the causative agent for blastomycosis, an endemic fungal infection prevalent in the Ohio and Mississippi River Valleys, Great Lakes region, and the Southeastern United States. It causes acute and chronic pneumonia, as well as disseminated extrapulmonary disease.
ABOUT THE KIT
This kit is for the quantitative determination of Blastomyces dermatitidis serum antigen in a human serum specimen. The Gotham Biotech Blastomyces dermatitidis Serum Antigen Detection Kit is a Research Use Only device for the determination of Blastomycosis in human patients as well as for monitoring of the efficacy of treatment.
SUMMARY AND EXPLANATION
Blastomycosis is a fungal infection in humans. The infection results in the release of antigen that may be detected in human serum. This assay employs antibodies specific to the Blastomyces dermatitidis antigen for the capture and detection of its presence in a human serum specimen. The detection method involves an enzyme/substrate system with the level of antigen in the serum proportional to the assay signal. The patient specimen result is compared to a standard curve of a series of assay calibrators to determine if the patient is positive or negative for Blastomycosis, and if positive, to determine the level of Blastomyces dermatitidis serum antigen in the specimen.
MEASUREMENT PRINCIPLE
The colorimetric reaction resulting from the detection of Blastomyces dermatitidis serum antigen in the specimen is measured on a spectrophotometer as absorbance at 450 nm, with a 650 nm filter blank.
REAGENTS
Allow all reagents to reach room temperature prior to use Assay Plate: 96 well microplate coated with Blastomyces dermatitidis specific antibody, and blocked. Store at 4- 8⁰ C. One plate included per kit 5X Wash Solution: A 5 fold concentrated wash solution employed in the wash steps of the assay. Store stock at room temperature.. Prepare working wash solution by adding 1 part 5X Wash Solution to 4 parts distilled water. 50 mL 5X stock included per kit. Working solution can be stored at 4- 8⁰ C for a period of 2 weeks.
Anti-Blastomyces dermatitidis-HRP Conjugate: This reagent, supplied as a 1000X stock is employed in the antigen detection step of the assay. Store stock at 4- 8⁰ C. The working stock is prepared by adding 1 part anti-Blastomyces dermatitidis-HRP conjugate stock to 999 parts Assay Conjugate Diluent. 20 μL 1000X stock is supplied with each kit. Prepare a fresh working stock for each assay.
Assay Conjugate Diluent: The assay conjugate diluent is used to dilute the 1000X anti-Blastomyces dermatitidis-
HRP conjugate. Store at 4- 8⁰ C. 30 mL of Assay Conjugate Diluent is included per kit
TMB Substrate: This reagent is the colorimetric substrate used in the development step of the assay. Store at 4- 8⁰ C. 30 ml of TMB substrate is included per kit.
Stop Solution: This reagent used to arrest the colorimetric reaction in the development step of the assay. Store at
4- 8⁰ C. 30 ml of TMB substrate is included per kit.
Materials and equipment required but not supplied:
Pipette (p200) and pipette tips
Multichannel pipette
Reagent reservoirs
Assay Plate Sealants
Clinical Laboratory Grade Distilled Water
Volumetric glassware or graduated cylinders
Timer
Liquid storage containers and test tubes (15 mL)
Vortex
DETERMINATIONS PER KIT
It is recommended that the calibrators, controls and patient serum specimens be run in duplicate. The Blastomyces dermatitidis Serum Antigen Detection Kit contains microwells and reagents sufficient to test 40 patient serum specimens run in duplicate in one single run, or 24 patient specimens run in duplicate for three runs, accounting for the duplicate runs of each calibrator and control.