Molecular discrimination of regulated and non-regulated Salmonella serotypes

Patent metadata

Field	Value
Publication number	US-10287638-B2
Application number	US-201515129108-A
Country	US
Kind code	B2
Filing date	Mar 26, 2015
Priority date	Mar 26, 2014
Publication date	May 14, 2019
Grant date	May 14, 2019

Abstract

The present invention provides a set of oligonucleotides to screen for the presence of targeted Salmonella serotypes in an enrichment or to characterize presumptive colonies. The set of oligonucleotides includes at least one set of primers and probe for the detection of Salmonella serotype selected from typhimurium, enteritidis, newport, heidelberg, infantis, virchow and Hadar . The set of oligonucleotides may include up to 5 different primer sets and the corresponding probes.

First claim

The invention claimed is:

1. A method of checking the safety of a foodstuff for Salmonella comprising the steps of:
providing a foodstuff suspected of being contaminated with Salmonella;
homogenizing the foodstuff;
extracting a nucleic acid sample from a homogenized foodstuff;
adding a set of primers to the nucleic acid sample, wherein the set of primers comprise at least one primer set selected from SEQ ID NOS: 1 and 3; SEQ ID NOS: 4 and 6; SEQ ID NOS: 7 and 9; and SEQ ID NOS: 10 and 12;
adding one or more Salmonella isolate identification probes to the nucleic acid sample, wherein the one or more Salmonella isolate identification probes are selected from SEQ ID NO: 2, SEQ ID NO: 5, SEQ ID NO: 8, and SEQ ID NO: 11;
amplifying the nucleic acid sample using the set of primers to from an amplified nucleic acid sample to form a double stranded DNA;
destroying the one or more Salmonella isolate identification probes as the double stranded DNA is formed;
releasing a fluorophore attached to the one or more Salmonella isolate identification probes; and
detecting the presence of the fluorophore, wherein a positive detection is indicative of at least one Salmonella being present in the sample.

2. The method of claim 1 , wherein the foodstuff is for human consumption, animal consumption or both.

3. The method of claim 1 , wherein the foodstuff is a food or a beverage.

4. The method of claim 1 , wherein the Salmonella is selected from Typhimurium, Enteritidis, Newport, Heidelberg, Infantis, Virchow and Hadar.

5. The method of claim 1 , wherein the foodstuff is selected from the group consisting of meat, poultry, pasteurized egg, and catfish products.

Assignees

• Univ Texas Tech System

Inventors

• Loneragan Guy
• Brashears Mindy M
• Nightingale Kendra
• Bugarel Marie

Classifications

• C12Q1/689 Primary

  for bacteria · CPC title

• Y02A50/451

  Cross-Sectional Technologies · mapped topic

• G01N33/02

  Food · CPC title

• C12Q2600/16

  Primer sets for multiplex assays · CPC title

• Y02A50/30

  Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change · CPC title