Novel diagnostic system for differentiation of enterobacteriaceae

Claim 2

A CULTURE MEDIUM HAVING A PH IN THE RANGE OF ABOUT 6.7-7.2 SUITABLE FOR DETERMINING THE IDENTIFICATION OF BACTERIA OF THE FAMILY ENTEROBACTERIACEAE WHICH COMPRISES A CHROMOGENIC B-GALACTOSIDASE SUBSTRATE IN COMBINATION WITH A MEMBER OF THE GROUP CONSISTING OF (A) A DECARBOXYLASE SUBSTRATE, (B) A DEAMINASE SUBSTRATE, (C) A UREASE SUBSTRATE, (D) A HYDROGEN SULFIDE DETECTING SYSTEM, OR (E) A CARBOHYDRATE FERMENTATION SYSTEM.

Claim 3

A culture medium having a pH in the range of about 6.7-7.2 suitable for determining the identification of bacteria of the family Enterobacteriaceae which comprises a chromogenic Beta -galactosidase substrate in combination with a member of the group consisting of (a) a decarboxylase substrate, (b) a deaminase substrate, (c) a urease substrate, (d) a hydrogen sulfide detecting system, or (e) a carbohydrate fermentation system.

Claim 4

A culture medium having a pH in the range of about 6.7-7.2 suitable for determining the identification of bacteria of the family Enterobacteriaceae which comprises a chromogenic Beta -galactosidase substrate in combination with a member of the group consisting of (a) a decarboxylase substrate, (b) a deaminase substrate, (c) a urease substrate, (d) a hydrogen sulfide detecting system, or (e) a carbohydrate fermentation system.

Claim 5

A culture medium of claim 1 wherein the chromogenic Beta -galactosidase substrate is chosen from the group consisting of o-nitrophenyl- Beta -galactopyranoside, 5-bromo-6-chloro-3-indolyl-Beta -D-galactoside and 6-bromo-2-naphthyl- Beta -D-galactoside, the decarboxylase agents are lysine, ornithine and arginine, the urease substrate is urea and a carbohydrate fermentation system selected from the group consisting of dextrose, mannitol, arabinose, sucrose, dulcitol, rhamnose, the deaminase substrate is selected from the group consisting or 1-aminoacids, preferably tryptophane, phenylalanine, and histidine, and the hydrogen sulfide detection system containing an iron salt in combination with hydrogen sulfide detecting agent preferably in the form of a thiosulfate.

Claim 6

A culture medium of claim 1 wherein the chromogenic Beta -galactasidase substrate is in combination with a decarboxylase substrate.

Claim 7

A culture medium of claim 1 wherein the chromogenic Beta -galactosidase substrate is in combination with a deaminase substrate.

Claim 8

A culture medium of claim 1 wherein the chromogenic Beta -galactosidase substrate is in combination with a hydrogen sulfide detecting system.

Claim 9

A culture medium of claim 1 wherein the chromogenic Beta -galactosidase substrate is in combination with a urease substrate.

Claim 10

A culture medium of claim 1 wherein the chromogenic Beta -galactosidase substrate is in combination with a carbohydrate fermentation system.

Claim 11

A culture medium having a pH in the range of about 6.7-7.2 suitable for determining the identification of bacteria of the Enterobacteriaceae which comprises the following ingredients, said ingredients being present in proportions indicated:

Claim 12

A culture medium having a pH in the range of about 6.7-7.2 suitable for determining the identification of bacteria of the Enterobacteriaceae which comprises the following ingredients, said ingredients being present in proportions indicated:

Claim 13

A process for preparing compositions of claim 8 which comprises admixing the bromthymol blue, yeast extract, dextrose, lysine, ferric ammonium citrate, sodium thiosulfate, agar, lactose and sodium chloride ingredients in suitable quantities of water, adjusting the pH to about 7.0, autoclaving the resulting solution followed by adding sterile o-nitrophenyl- Beta -galactopyranoside, urea and tryptophan, with q.s. water to form the defined concentrations.

Claim 14

The process for the identification of bacteria of the Enterobacteriaceae which comprises inoculating the media defined by claim 8 allowing the inoculated media to grow for at least about 24 hours at 37*C, followed by the identification of the bacteria according to the herein described changes for lysine, hydrogen sulfide, tryptophan, ONPG, urea and indole.

Claim 15

The process for the identification of bacteria of the Enterobacteriaceae which comprises inoculating the media defined by claim 8 allowing the inoculated media to grow for at least about 24 hours at 37*C, followed b the idenification of the bacteria according to the herein described color changed for glucose, lysine, hydrogen sulfide, tryptophan, o-nitrophenyl Beta -galactopyranoside urea and indole.

Claim 16

A sterile culure medium having a pH of about 6.7-7.2 comprising:

Claim 17

A sterile culure medium having a pH of about 6.7-7.2 comprising:

Claim 18

A process for preparing composition of claim 12, which comprises admixing the bromthymol blue, yeast extract, dextrose, lysine, ferric ammonium citrate, sodium thiosulfate, agar, lactose and sodium chloride ingredients in suitable quantities of water, adjusting the pH to about 7.0, autoclaving the resulting solution followed by adding sterile o-nitrophenyl- Beta -galactopyranoside urea and tryptophan, with q.s. water to form the defined concentrations.

Claim 19

The process for the identification of bacteria of the Enterobacteriaceae which comprises inoculating the media defined by claim 12, allowing the inoculated media to grow for at least about 24 hours at 37*C, followed by the identification of the bacteria according to the herein described changes for lysine, hydrogen sulfide, tryptophan, ONPG, urea and indole.

Claim 20

The process of claim 14 whErein the color change results are read according to the following table:

Claim 21

The process of claim 14 wherein the color change results are read according to the following chart:

Claim 22

The process for the identification of bacteria of the Enterobacteriaceae which comprises inoculating the media defined by claim 12, allowing the inoculated media to grow for at least about 24 hours at 37*C, followed by the identification of the bacteria according to the herein described color changes for glucose, lysine, hydrogen sulfide, tryptophan, o-nitrophenyl Beta -galactopyranoside urea and indole.