# US 9,518,283 B1 · Selective enrichment media and uses thereof

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- Number: US 9,518,283 B1
- Document ID: US9518283B1
- Status: Granted
- Kind: B1
- Application number: 14/189,488
- Filed: 2014-02-25
- Granted: 2016-12-13
- Earliest claimed: 2012-02-27
- Assignees: Paradigm Diagnostics, Inc.
- Applicants: Paradigm Diagnostics, Inc.
- Inventors: Alan D. Olstein
- Web page: https://patentgrep.com/patent/US9518283B1

## CPC

- C12Q 1/10: Enterobacteria
- C12N 1/20: Bacteria; Culture media therefor
- C12N 1/38: Chemical stimulation of growth or activity by addition of chemical compounds which are not essential growth factors; Stimulation of growth by removal of a chemical compound (C12N1/34 takes precedence)
- C12Q 1/045: {Culture media therefor}
- G01N 2333/245: Escherichia (G)
- G01N 2333/255: Salmonella (G)
- Y02A 50/30: Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)
- Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE

## Abstract

Selective enrichment media and methods for selectively growing and detecting Salmonella spp. and/or Shiga toxin-producing E. coli. The media may comprise a carbon and nitrogen source, an inorganic salt, a fermentable sugar, one or more selective agents, and an efflux pump inhibitor. Various selective agents include sulfa drugs, surfactants, aminocoumarins, cycloheximide, supravital stains, ascorbic acid, bromobenzoic acid, myricetin, nitrofurantoin, rifamycins, polyketides, and oxazolidinones. Various efflux pump inhibitors include arylpiperazines, such as 1-(1-naphthylmethyl)piperazine, and quinoline derivatives, such as 4-chloroquinoline. Methods of selectively growing and detecting Salmonella and/or Shiga toxin-producing E. coli are provided.

## Claims

### Claim 1

A selective enrichment medium for Salmonella or Shiga toxin-producing E. coli comprising:
a carbon and nitrogen source;
an inorganic salt;
a selective agent comprising one or more of a sulfa drug, a surfactant, an aminocoumarin, cycloheximide, myricetin, and nitrofurantoin and one or more of a rifamycin, a polyketide, and an oxazolidinone; and
an efflux pump inhibitor comprising one or more of 1-(1-naphthylmethyl)piperazine and 4-chloroquinoline.

### Claim 2 (depends on 1)

The selective enrichment medium of claim 1 wherein the efflux pump inhibitor comprises 1-(1-naphthylmethyl)piperazine.

### Claim 3 (depends on 1)

The selective enrichment medium of claim 1 wherein the efflux pump inhibitor comprises 4-chloroquinoline.

### Claim 4 (depends on 1)

The selective enrichment medium of claim 1 wherein the selective agent comprises a sulfa drug, wherein the sulfa drug comprises one or more of sulfanilamide and sulfathiazole.

### Claim 5 (depends on 1)

The selective enrichment medium of claim 1 further comprising one or more of a fermentable sugar, a supravital stain, ascorbic acid, and bromobenzoic acid, wherein the fermentable sugar comprises 2-deoxy-D-ribose and the supravital stain comprises brilliant green.

### Claim 6 (depends on 1)

The selective enrichment medium of claim 1 wherein the selective agent comprises: a surfactant, wherein the surfactant comprises 7-ethyl-2-methyl-4-undecyl sulfate or a salt thereof; and an aminocoumarin, wherein the aminocoumarin comprises novobiocin.

### Claim 7 (depends on 1)

The selective enrichment medium of claim 1 wherein the selective agent comprises a rifamycin, wherein the rifamycin comprises rifampicin.

### Claim 8 (depends on 1)

The selective enrichment medium of claim 1 wherein the selective agent comprises a polyketide, wherein the polyketide comprises doxycycline.

### Claim 9 (depends on 1)

The selective enrichment medium of claim 1 wherein the selective agent comprises an oxazolidinone, wherein the oxazolidinone comprises linezolid.

### Claim 10 (depends on 1)

The selective enrichment medium of claim 1 wherein the selective agent comprises one or more of rifampicin, tetracycline, doxycycline, and linezolid.

### Claim 11 (depends on 1)

The selective enrichment medium of claim 1 wherein the selective agent comprises a sulfa drug, a surfactant, an aminocoumarin, cycloheximide, myricetin, nitrofurantoin, and one or more of a rifamycin, a polyketide, and an oxazolidinone.

### Claim 12 (depends on 1)

The selective enrichment medium of claim 1 further comprising a visual indicator that indicates the presence of a microorganism selected from the group consisting of Salmonella and E. coli.

### Claim 13 (depends on 1)

The selective enrichment medium of claim 1 wherein the selective agent and the efflux pump inhibitor are present in amounts effective to inhibit growth of at least one non-Salmonella species to a greater extent than one or more Salmonella species.

### Claim 14

A selective enrichment medium for Salmonella or Shiga toxin-producing E. coli comprising:
a carbon and nitrogen source;
an inorganic salt;
a selective agent comprising one or more of a sulfa drug, a surfactant, an aminocoumarin, cycloheximide, myricetin, nitrofurantoin, a rifamycin, a polyketide, and an oxazolidinone; and
an efflux pump inhibitor comprising one or more of 1-(1-naphthylmethyl)piperazine and 4-chloroquinoline,
wherein the selective agent and the efflux pump inhibitor are present in amounts effective to inhibit growth of at least one non-Shiga toxin-producing E. coli strain to a greater extent than one or more Shiga toxin-producing E. coli strains.

### Claim 15 (depends on 1)

A method of selectively enriching for a microorganism selected from the group consisting of a Salmonella species and a Shiga toxin-producing E. coli strain, comprising culturing a sample suspected of containing the microorganism in the selective enrichment medium of claim 1.

### Claim 16 (depends on 15)

The method of claim 15 wherein the culturing grows a Salmonella species, a Shiga toxin-producing E. coli strain, or both a Salmonella species and a Shiga toxin-producing E. coli strain.

### Claim 17 (depends on 16)

The method of claim 16 further comprising detecting presence of the microorganism.

### Claim 18 (depends on 1)

The selective enrichment medium of claim 1 wherein the selective agent and the efflux pump inhibitor are present in amounts effective to inhibit growth of at least one non-Shiga toxin-producing E. coli strain to a greater extent than one or more Shiga toxin-producing E. coli strains.