Appeal No. 1999-1206 Page 2
Application No. 08/394,608
Claims 18, 22, 28, 33 and 36 are representative of the subject matter on appeal
and read as follows:
18. A test medium for detecting the presence of biological material having
β-galactosidase enzyme specificity, said test medium comprising:
a base medium for maintenance of said biological material; and
a chromogenic indolyl β-galactoside substrate forming an insoluble compound of
a first color upon reacting with β-galactosidase, wherein said chromogenic
β-galactoside substrate is selected from the group consisting of 6-chloroindolyl-β-D-
galactoside, 4,6-dichloroindolyl-β-D-galactoside, 6, 7-dichloroindolyl-β-D-galactoside,
4,6, 7-trichloroindolyl-β-D-galactoside and salts thereof.
22. The test medium of claim 18, further comprising 5-bromo-4-chloro-3-indolyl-
β-glucuronide.
28. A method for testing the presence of and quantitatively identifying and
differentiating microorganisms in a biological test sample, said test sample containing
first microorganisms having β-galactosidase but not β-glucuronidase activity, and
second microorganisms having β-glucuronidase activity comprising the steps of
inoculating a solid test medium or substrate capable of forming a solid with said
test sample, said test medium comprising a chromogenic β-galactoside substrate
capable of forming a first water-insoluble precipitate of a first color upon reacting with β-
galactosidase, a chromogenic β-glucuronide substrate capable of forming a second
water-insoluble precipitate of a second color contrasting with said first color upon
reacting with β-glucuronidase, and a base medium, said first and second colors being
visibly distinguishable in daylight,
incubating said test medium to produce colonies of said first and second
microorganisms and first and second colored precipitates corresponding to said
colonies,
examining said test medium for the presence of colonies having said first color,
such colonies being colonies of microorganisms having β-galactosidase but not β-
glucuronidase activity, and the presence of colonies having said second color, such
colonies being colonies of microorganisms having β-glucuronidase activity, and
enumerating said colonies of microorganisms having β-galactosidase activity and
said colonies of microorganisms having β-glucuronidase activity.
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