IP Library Granted Patent US 10,519,481
Granted Patent B2
US 10,519,481 · App. 15/593,598 · Granted Dec 31, 2019

Method of detecting a

Inventors: Christine A. Binsfeld (Woodbury, MN); Patrick A. Mach (Shorewood, MN); Mara S. Celt (Red Wing, MN); Adam J. Stanenas (Cottage Grove, MN)
Assignee: 3M Innovative Properties Company
C12Q1/10C12Q1/04C12Q1/34C12Y302/01023C12Q2304/00C12Q2334/00
View Patent ↗
Loading inventors, assignments & file history…
Monitor This Case
Get email alerts when status or documents change.
Order Certified Copies
Most orders are placed with the USPTO same day — all within 24 business hours.
Order via The Patent Place →
Pre-filled with this patent's details
Quick Facts
Patent No.
US 10,519,481
App. No.
15/593,598
Granted
Dec 31, 2019
Kind
B2
Abstract

A method of detecting a Salmonella microorganism is provided. The method includes the use of a selective growth medium, a first indicator system that is converted to a first detectable product by a Salmonella microorganism, and a second indicator system that is converted to a second detectable product by β-galactosidase enzyme activity. The method further comprises inoculating the growth medium and incubating the inoculated growth medium at a temperature higher than 40 degrees C.

Claims (36)

1. A method of detecting a microorganism, the method comprising: providing,

a sample to be tested;

a culture device;

a nutrient medium that facilitates growth of a Gram-negative enteric microorganism;

a first selective agent that inhibits the growth of Gram-positive microorganisms;

a first differential indicator system comprising a first differential indicator compound that can be converted to a first detectable product by a member of a group of Salmonella microorganisms that includes a microorganism of the species Salmonella bongori;

a second differential indicator system comprising a second differential indicator compound that can be converted to a second detectable product by a β-galactosidase enzyme activity; and

a third differential indicator system comprising a third differential indicator compound that can be converted by a Salmonella microorganism to a third detectable product;

contacting in the culture device the nutrient medium, the first selective agent, the first differential indicator system, and the second differential indicator system with a sample to form an inoculated culture device;

incubating the inoculated culture device for a first period of time at a temperature greater than 40 degrees C.;

detecting the presence of the first detectable product;

detecting the absence of the second detectable product; and

detecting the presence or absence third detectable product;

wherein observing the presence of the third detectable product indicates the presence of a Salmonella microorganism in the sample; and

wherein observing the absence of the third detectable product indicates the absence of a Salmonella microorganism in the sample.

2. The method of claim 1 , wherein observing the absence of the third detectable product indicates the presence of a non- Salmonella microorganism in the sample.

3. The method of claim 1 , wherein detecting the the third detectable product indicates the presence of a Salmonella microorganism that is selected from the group consisting of S. enterica, S. bongori , and combinations thereof.

4. The method of claim 3 , wherein detecting the presence of the third detectable product indicates the presence of a Salmonella microorganism of the species S. bongori.

5. The method of claim 1 , further comprising enumerating a number of colonies formed by microorganisms belonging to a group consisting of Salmonella microorganisms and β-galactosidase-producing microorganisms.

6. The method of claim 1 , wherein observing the culture device comprises observing the culture device visually.

7. The method of claim 1 , wherein observing the culture device comprises creating an image of the culture device using an imaging device.

8. The method of claim 7 , further comprising analyzing the image using a processor.

9. The method of claim 1 , wherein the culture device is provided as a thin film culture device with the nutrient medium, the first selective agent, the first differential indicator system, and the second differential indicator system disposed therein in a dehydrated form.

10. The method of claim 1 , wherein the first differential indicator system comprises a pH indicator and at least one carbohydrate selected from the group consisting of melibiose, 2-deoxy-D-ribose, mannitol, L-arabinose, dulcitol, maltose, L-rhamnose, trehalose, D-xylose, and sorbitol.

11. The method of claim 1 , wherein the second differential indicator compound is selected from the group consisting of 5-bromo-4-chloro-3-indolyl-β-D-galactopyranoside, 5-bromo-3-indolyl-β-D-galactopyranoside, 5-bromo-6-chloro-3-indolyl-β-D-galactopyranoside, 2-nitrophenyl-β-D-galactopyranoside, and 4-nitrophenyl-β-D-galactopyranoside.

12. The method of claim 1 , wherein the first selective agent is selected from the group consisting of an antibiotic, bile salts, bile salts No. 3, deoxycholic acid, cholic acid, crystal violet, novobiocin, nalidixic acid, polymyxin B, streptomycin, methicillin, cefsoludin, or a combination of any two or more of the foregoing selective agents.

13. The method of claim 1 , wherein the culture device further comprises at least one second selective agent, wherein the at least one second selective agent inhibits the growth of at least one Gram-negative enteric microorganism that is not a member of the genus Salmonella.

14. The method of claim 13 , wherein the at least one second selective agent comprises a combination of nalidixic acid, streptomycin, and polymyxin B.

15. The method of claim 1 , further comprising providing a nondifferential indicator compound that can be converted by an enteric microorganism to a fourth detectable product; and

observing the culture device to detect a presence or an absence of the fourth detectable product;

wherein contacting in the culture device the nutrient medium, the first selective agent, the first differential indicator system, and the second differential indicator system with a sample to form an inoculated culture device further comprises contacting in the culture device the nutrient medium, the first selective agent, the first-differential indicator system, the second differential indicator system, and the nondifferential indicator compound with the sample to form the inoculated culture device.

16. The method of claim 1 , wherein detecting the presence of the first detectable product comprises observing the culture device to detect a first detectable color.

17. The method of claim 1 , wherein the third differential indicator system comprises a pH indicator and a nutrient, a chromogenic enzyme substrate, or a combination thereof.

18. The method of claim 1 , wherein the third detectable product is optically distinguishable from the first detectable product and the second detectable product.

19. The method of claim 1 , wherein the first differential indicator system comprises an enzyme substrate.

20. The method of claim 19 , wherein the enzyme substrate comprises a substance that is selected from the group consisting of 5-bromo-6-chloro-3-indolyl caprylate, 4-nitrophenyl caprylate, 2-naphthyl caprylate, 5-bromo-4-chloro-3-indoxyl-α-D-galactopyranoside, resorufinyl-α-D-galactopyranoside, and 4-nitrophenyl-α-D-galactopyranoside to detect caprylate esterase enzyme activity.

Assignments (3)
SECURITY INTEREST Recorded Sep 2, 2022
From: NEOGEN CORPORATION; NEOGEN FOOD SAFETY US HOLDCO CORPORATION
To: JPMORGAN CHASE BANK, N.A., AS COLLATERAL AGENT
Reel/Frame 061372/0264 →
PATENT ASSIGNMENT AGREEMENT Recorded Sep 1, 2022
From: 3M INNOVATIVE PROPERTIES COMPANY
To: GARDEN US HOLDCO CORPORATION
Reel/Frame 061365/0688 →
CHANGE OF NAME Recorded Sep 1, 2022
From: GARDEN US HOLDCO CORPORATION
To: NEOGEN FOOD SAFETY US HOLDCO CORPORATION
Reel/Frame 061370/0437 →
Continuity (3)
Continuation 14368525
Provisional Application 61580860 · Dec 28, 2011
Related Publication 20170247738A1 · Aug 31, 2017