Specific oligonucleotide primers for detection of pathogenic campylobacter bacteria by polymerase chain reaction

• US 5,494,795 A
• Filed: 05/05/1993
• Issued: 02/27/1996
• Est. Priority Date: 05/05/1993
• Status: Expired due to Fees

• Current Assignee: the united states of america as represented by the secretary of the navy
• Original Assignee: the united states of america as represented by the secretary of the navy
• Inventors: Guerry, Patricia, Trust, Trevor J.
• Primary Examiner(s): Zitomer, Stephanie W.
• Assistant Examiner(s): MYERS, CARLA J
• Application Number:
  US08/056,538
• Time in Patent Office: 1,028 Days
• Field of Search: 536/23.7, 536/24.32, 536/24.33, 435/91, 435/6, 435/91.2, 935/78, 514/44
• US Class Current: 435/6.11
• CPC Class Codes: C12Q 1/689 for bacteria

Abstract

This invention is a specific set of oligonucleotide PCR primers (pg50 and pg3) and a specific oligonucleotide probe (pBA273) for detection of PCR-amplified DNA from pathogenic strains of Campylobacter, specifically C. coli and C. jejuni, in fecal specimens.

Claims

1. An oligonucleotide 5'"'"'-ATGGGATTTCGTATTAAC-3'"'"'(SEQ ID No. 1).

2. An oligonucleotide 5'"'"'-GAACTTGAACCGATTTG-3'"'"'(SEQ ID No. 2).

3. A composition comprising a primer pair as used for amplifying DNA of the flagellin gene by polymerase chain reaction (PCR) from pathogenic strains Campylobacter coli and Campylobacter jejuni of Campylobacter wherein the primer pair consists of 5'"'"'-ATGGGATTTCGTATTAAC-3'"'"'(SEQ ID No. 1) and 5'"'"'-GAACTTGAACCGATTTG-3'"'"'(SEQ) ID No. 2).

4. The composition of claim 3 in combination with a labeled probe detecting sequences internal to the DNA amplified by the primer pair.

5. The composition of claim 4 wherein the probe is a segment of the Campylobacter flagell in gene.

6. A method of detecting the pathogenic strains Campylobacter coli and Campylobacter jejuni in stool specimens of patients with idiopathic diarrhea by PCR comprising amplifying the pathogenic strains with a primer pair consisting of 5'"'"'-ATGGGATTTCGTATTAAC-3'"'"'(SEQ ID No. 1) and 5'"'"'-GAACTTGAACCGATTTG-3'"'"'(SEQ ID No. 2) and detecting the amplified DNA by observation of the amplified DNA fragment.