# US 2013/0280714 A1 · Rapid salmonella serotyping assay

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- Number: US 2013/0280714 A1
- Document ID: US20130280714A1
- Status: Published application
- Kind: A1
- Application number: 13/871,098
- Filed: 2013-04-26
- Published: 2013-10-24
- Earliest claimed: 2010-10-26
- Assignees: Centers for Disease Control and Prevention
- Applicants: Centers for Disease Control and Prevention
- Inventors: Patricia Fields; John R. McQuiston; Collette Fitzgerald Leaumont
- Web page: https://patentgrep.com/patent/US20130280714A1

## CPC

- C12Q 1/689: for bacteria
- C12Q 2600/16: Primer sets for multiplex assays
- 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
- 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

Processes for the serotype specific detection and identification of one or more Salmonella serotypes are provided. A family of specific primers and probes are provided that allow screening of biological or environmental samples for robust, rapid, and reproducible detection and identification of one or more Salmonella serotypes in the sample.

## Claims

### Claim 1

A process of serotyping Salmonella in a sample comprising:
adding to a sample one or more O-grp-2 primer sets operable to produce an O-grp-2 amplification product of an O-grp-2 target sequence;
said primer set comprising a forward primer and a reverse primer that hybridize to distinct regions flanking said O-grp-2 target sequence under conditions suitable for a polymerase chain reaction; and
detecting the presence or absence of said O-grp-2 amplification product by hybridization of one or more labeled probes specific to said O-grp-2 amplification product, said labeled probe comprising a label and a nucleotide sequence, said nucleotide sequence comprising the sequence of SEQ ID NO: 131, 132, 133, 134 or 135.

### Claim 2 (depends on 1)

The process of claim 1 further comprising detecting the presence or absence of one or more O-grp-1 amplification products, one or more H-ag amplification products, or both.

### Claim 3 (depends on 1)

The process of claim 1 wherein said detecting the presence or absence of said O-grp-2 amplification product is performed in the same sample with detection of one or more O-grp-1 amplification products, one or more H-ag amplification products, or both.

### Claim 4 (depends on 1)

The process of claim 1 wherein at least one of said forward primers comprises the nucleotide sequence of any one or more of SEQ ID NOs: 83, 84, 85, 92, 93, 94, 95, 96, 107, 109, 111, 113, or 115.

### Claim 5 (depends on 1)

The process of claim 1 wherein at least one of said reverse primers comprises the nucleotide sequence of any one or more of SEQ ID NOs: 86, 87, 88, 97, 98, 99, 100, 101, 108, 110, 112, 114, or 116.

### Claim 6 (depends on 1)

The process of claim 1 wherein said nucleotide sequence consists of the sequence of SEQ ID NO: 131, 132, 133, 134, or 135.

### Claim 7 (depends on 1)

The process of claim 1 wherein said detecting diagnoses Salmonella infection in a subject.

### Claim 8

A probe for detection of a nucleotide sequence in a sample comprising
an isolated nucleotide sequence consisting of the sequence of SEQ ID NO: 131, 132, 133, 134 or 135; and
a label.

### Claim 9 (depends on 8)

The probe of claim 8 wherein said label is a particle.

### Claim 10

A kit for serotyping Salmonella in a sample comprising:
a probe of 20 to 40 nucleotides, said probe capable of hybridizing to an O-grp-2 amplicon.

### Claim 11 (depends on 10)

The kit of claim 10 wherein
said O-grp-2 probe comprises a nucleotide sequence, said nucleotide sequence comprising SEQ ID NOs: 131, 132, 133, 134 or 135.

### Claim 12 (depends on 10)

The kit of claim 10 further comprising
a plurality of O-grp-1 primers,
a plurality of H-ag primers,
a plurality of O-grp-1 probes, and
a plurality of H-ag probes.

### Claim 13 (depends on 12)

The kit of claim 12 wherein
said O-grp-1 primers comprise at least one of those of Table 1;
said H-ag primers comprise at least one of those of Table 2;
said O-grp-1 probes comprise at least one of those of Table 1; or
said H-ag probes comprise at least one of those of Table 2.

### Claim 14 (depends on 10)

The kit of claim 10 wherein said nucleotide sequence consists of a sequence of SEQ ID NOs: 131, 132, 133, 134 or 135.

### Claim 15 (depends on 10)

The kit of claim 10 wherein said probe further comprises a label.

### Claim 16 (depends on 15)

The kit of claim 15 wherein said label is a particle.