Method for detecting staphylococci

Abstract

The present invention exploits the herein first reported, empirical observation that even though staphylococci produce the enzyme, beta-glucosidase, this genus of bacteria is not able to produce a metabolite that will enzymatically react with 5-bromo-4-chloro-3-indolyl-beta-D-glucopyranoside, a substrate commonly used to detect the presence of beta-glucosidase produced by other bacteria. In view of this unexpected observation, one embodiment of the present invention includes a selective medium containing inhibitors to enhance staphylococci growth as well as a first glucopyranoside substrate, such as 5-bromo-4-chloro-3-indolyl-beta-D-glucopyranoside, and a second phosphatase substrate, such as 6-chloro-3-indolylphosphate or 5-bromo-6-chloro-3-indolylphosphate. In this embodiment, staphylococci in a sample will produce metabolites that will react with the phosphate substrate in the medium to produce colonies having red to red-violet color while other bacteria in the sample will produce beta-glucosidase that will react with the glucopyranosidase substrate in the medium and produce colonies having a blue color. In a particularly preferred embodiment of this invention, a thin film culture plate device, such as a PETRIFILM culture plate device, is prepared using a dry culture medium containing selected inhibitors, a first glucopyranoside substrate and a second phosphate substrate. When the thin film culture plate device is inoculated with a sample and then incubated for a sufficient period of time, staphylococci in the sample will produce colonies on the thin film device having a red color and other bacteria in the sample will produce colonies on the device having a blue color.