Culture medium for rapid count of coliform bacteria

• US 5,364,766 A
• Filed: 05/14/1993
• Issued: 11/15/1994
• Est. Priority Date: 05/14/1993
• Status: Expired due to Term

• Current Assignee: 3M Company
• Original Assignee: 3M Company
• Inventors: Mach, Patrick A., Adams, Carl A., Hesselroth, Karen E., Schwab, Debra L.
• Primary Examiner(s): Wityshyn, Michael G.
• Assistant Examiner(s): Mohamed, Abdel A.
• Application Number:
  US08/062,311
• Time in Patent Office: 550 Days
• Field of Search: 435/34.4, 435/29, 435/38, 435/39, 435/240.3, 435/240.31, 435/243, 435/252.33, 435/261, 435/810, 435/849, 422/55, 422/56, 422/57, 422/61
• US Class Current: 435/34
• CPC Class Codes: C12M 23/04 Flat or tray type, drawers C12M23/10, C12M23/12, C12M23/16 take precedence
• CPC Class Codes: C12M 23/22 Transparent or translucent parts glassware for laboratory use B01L3/00
• CPC Class Codes: C12M 23/26 flexible flexible containers for laboratory use B01L3/505
• CPC Class Codes: C12M 23/38 Caps; Covers; Plugs; Pouring means
• CPC Class Codes: C12M 25/06 Plates; Walls; Drawers; Multilayer plates
• CPC Class Codes: C12M 41/36 of biomass, e.g. colony counters or by turbidity measurements electrooptical investigation of individual particles G01N15/14, flow cytometers G01N15/1404
• CPC Class Codes: C12N 1/20 Bacteria; Culture media therefor
• CPC Class Codes: C12Q 1/045 Culture media therefor
• CPC Class Codes: Y10S 435/81 Packaged device or kit
• CPC Class Codes: Y10S 435/849 Escherichia coli

Abstract

This invention generally relates to products and processes used to determine the presence of bacteria in a sample and particularly relates to a culture medium which may be used in products and processes to allow early detection and count of coliform bacteria. The bacterial culture medium which facilitates the early detection and count of coliform bacteria is a mixture of tryptose, lactose, sodium chloride, bile salts, guar gum and an excess amount of phenol red sufficient to provide a high concentration of phenol red in close proximity to the growing bacteria in order to allow detection and count of the growing bacteria in less than 12 hours.

Claims

1. A bacterial culture medium which facilitates the early detection and count of coliform bacteria growing in the medium comprising tryptose, lactose, sodium chloride, bile salts, guar gum and an excess amount of phenol red sufficient to provide a high concentration of phenol red in close proximity to the growing bacteria in order to allow detection and count of the growing bacteria in less than 12 hours.

2. The culture medium of claim 1 wherein the concentration of phenol red is in the range of about 160 mg/l to about 5000 mg/l.

3. The culture medium of claim 1 wherein the concentration of phenol red is in the range of about 1000 mg/l about 5000 mg/l.

4. The culture medium of claim 1 wherein the medium further comprises triphenyltetrazolium chloride.

5. The culture medium of claim 4 containing about 10-20 g/l tryptose, 2.5-7.5 g/l lactose, 2.5-7.5 g/l sodium chloride, 1.35-1.65 g/l bile salts, 2.5-7.5 g/l guar gum, 0.025-0.120 g/l triphenyltetrazolium chloride, and 0.16-5.0 g/l phenol red.

6. The culture medium of claim 1 containing about 15 g/l tryptose, 5 g/l lactose, 5 g/l sodium chloride, 1.5 g/l bile salts, 5 g/l guar gum, 0.050 g/l triphenyltetrazolium chloride and 1.25 g/l phenol red.

7. A method of detecting the presence of coliform bacteria in a sample comprising the steps of adding an aliquot of the sample containing coliform bacteria to a culture medium comprising tryptose, lactose, sodium chloride, bile salts, guar gum and an excess amount of phenol red sufficient to provide a high concentration of phenol red in close proximity to the bacteria;

growing the bacteria in the presence of the culture medium; and

detecting the color change of the phenol red as the growing bacteria metabolize in less than about 12 hours.

8. The method of claim 7 wherein the bacteria are grown in agar containing the culture medium.

9. The method of claim 7 wherein the bacteria are grown in a suspension containing the medium.

10. The method of claim 9 wherein the bacteria are grown in a thin film containing the medium using a device having a self-supporting, waterproof substrate and a transparent cover sheet, wherein the culture media is coated on the waterproof substrate and then dried.

11. The method of claim 7 wherein the color change of the phenol red is detected visually.

12. The method of claim 7 wherein the color change of the phenol red is detected by an instrument.

13. The method of claim 7 wherein the culture medium further comprises triphenyltetrazolium chloride.

14. The method of claim 13 further comprising the step confirming the presence of coliform bacteria in the sample by detecting gas formation and a color change in the triphenyltetrazolium chloride.

15. A device to detect coliform bacterial growth in a sample consisting essentially of a self-supporting, waterproof substrate, a foam spacer and a transparent cover sheet, wherein a culture medium comprising tryptose, lactose, sodium chloride, bile salts, guar gum and an excess amount of phenol red is coated on the self-supporting, waterproof substrate sufficient to provide a high concentration of phenol red in close proximity of growing bacteria in order to allow detection and count of the growing bacteria in less than 12 hours.

16. The device of claim 15 wherein the phenol red changes color from red to yellow and wherein the color change is detected visually.

17. The device of claim 15 wherein the phenol red changes color from red to yellow and wherein the color change is detected by an instrument.

18. The device of claim 15 wherein the culture medium further comprises triphenyltetrazolium chloride.

19. The device of claim 18 wherein the triphenyltetrazolium chloride changes color in the presence of coliform bacteria after about 24 hours.