Probes and methods for the detection of salmonella

• US 5,147,778 A
• Filed: 11/29/1988
• Issued: 09/15/1992
• Est. Priority Date: 12/01/1987
• Status: Expired due to Term

• Current Assignee: Amoco Corporation (BP PLC)
• Original Assignee: Amoco Corporation (BP PLC)
• Inventors: Wilson, Stephen G., Shah, Jyotsna, Lane, David J., Halbert, Donald N., Nietupski, Raymond M., Chan, Samuel W.
• Primary Examiner(s): Moskowitz, Margaret
• Assistant Examiner(s): Zitomer, Stephanie W.
• Application Number:
  US07/277,579
• Time in Patent Office: 1,386 Days
• Field of Search: 435/6, 435/810, 435/879, 436/501, 436/94, 536/28, 536/27, 935/77, 935/78
• US Class Current: 435/6.11
• CPC Class Codes: C12Q 1/6813 Hybridisation assays
• CPC Class Codes: C12Q 1/689 for bacteria
• CPC Class Codes: 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
• CPC Class Codes: Y10S 435/879 Salmonella
• CPC Class Codes: Y10T 436/143333 Saccharide [e.g., DNA, etc.]

Abstract

Improved nucleic acid probes capable of specifically hybridizing to rRNA of Salmonella and not to rRNA of non-Salmonella are described along with methods utilizing such probes for the detection of Salmonella in food and other samples.

Claims

1. A nucleic acid probe consisting essentially of a sequence of nucleotides of any one of probes 849, 1069, 1166, or 1200, or nucleotide sequences complementary thereto, which nucleic acid probe preferentially hybridizes to rRNA or rDNA of Salmonella over rRNA or rDNA of non-Salmonella bacteria.

2. A set of nucleic acid probes selected from the group of probe sets consisting of probes 1200 and 1165;

1200 and 1166;

1165 and 1166;

849 and 1166;

849 and 1069;

849, 1200 and 1166;

1165, 1200 and 1166;

849, 1165, 1200 and 1166;

each nucleic acid probe consisting essentially of a sequence of nucleotides of any one of said probes in said sets, or nucleotide sequences complementary thereto, which nucleic acid probes preferentially hybridize to rRNA or rDNA of Salmonella over rRNA or rDNA of non-Salmonella bacteria.

3. A method for detecting the presence of Salmonella in a sample comprising:

a) contacting said sample with at least one nucleic acid probe consisting essentially of a sequence of nucleotides of any one of probes 849, 1069, 1166, or 1200, or nucleotide sequences complementary thereto, which nucleic acid probe preferentially hybridizes to rRNA or rDNA of Salmonella over rRNA or rDNA of non-Salmonella bacteria;

b) imposing hybridization conditions on said sample and the nucleic acid probe to allow the nucleic acid probe to hybridize to the rRNA or rDNA of Salmonella, if present in said sample to form hybridized nucleic acid complexes, under conditions which do not allow said nucleic acid probe to form stable hybridization nucleic acid complexes with non-Salmonella bacteria rRNA or rDNA; and

c) detecting said hybridized nucleic acid complexes as an indication of the presence of said Salmonella in said sample.

4. The method of claim 3 wherein said nucleic acid probe consists essentially of any one of probes 1166, 1069, and 849 or nucleotide sequences complementary thereto.

5. A method for detecting Salmonella in a sample comprising the steps of:

a) contacting a sample under hybridization conditions with a set of nucleic acid probes selected from the group of probe sets consisting of probes 1200 and 1165;

1200 and 1166;

1165 and 1166;

849 and 1166;

849 and 1069;

849, 1200 and 1166;

1165, 1200 and 1166;

849, 1165, 1200 and 1166;

or nucleotide sequences complementary thereto, which nucleic acid probe preferentially hybridizes to rRNA or rDNA of Salmonella over rRNA or rDNA of non-Salmonella bacteria; and

b) detecting hybridized nucleic acid complexes as an indication of the presence of Salmonella in said sample.

6. An assay kit comprising a nucleic acid probe consisting essentially of a sequence of nucleotides of any one of probes 849, 1069, 1166, or 1200, or nucleotide sequences complementary thereto, which nucleic acid probe preferentially hybridizes to rRNA or rDNA of Salmonella over rRNA or rDNA of non-Salmonella bacteria.

7. A nucleic acid probe which preferentially hybridizes to rRNA or rDNA of Salmonella over rRNA or rDNA of non-Salmonella bacteria and consists essentially of the base sequence of probe 1200, or nucleotide sequences complementary thereto.

8. A nucleic acid probe which preferentially hybridizes to rRNA or rDNA of Salmonella over rRNA or rDNA of non-Salmonella bacteria and consists essentially of the base sequence of probe 849, or nucleotide sequences complementary thereto.