Methods and reagents for the detection of herpes simplex virus, treponema pallidum, and haemophilus ducreyi
AssigneeHoffmann-La Roche Inc.
InventorsKarina A. Orle, Judith B. Weiss
Life of this patent
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Abstract
The present invention provides methods and reagents for the detection and identification of four causative agents of genital ulcerations, herpes simplex virus (HSV) types 1 and 2, Treponema pallidum, and Haemophilus ducreyi. The methods use sequence-specific primers which enable the simultaneous polymeric chain reaction amplification of genomic nucleic acid sequences from HSV types 1 and 2, T. pallidum, and H. ducreyi. Following amplification, sequence-specific oligonucleotide probes are used to detect and distinguish HSV type 1 and 2, T. pallidum, and H. ducreyi nucleic acid.
Claims14
A pair of oligonucleotides primers for the amplification of nucleic acid from a region of the Treponema pallidum 47 kilodalton membrane immunogen gene, wherein said pair of primers is KO3A (SEQ ID NO:
- 6) and KO4 (SEQ ID NO:
- 7).
A pair of oligonucleotides primers for the amplification of nucleic acid from a region of the Haemophilus ducreyi 16S ribosomal RNA gene, wherein said pair of primers is KO7A (SEQ ID NO:
- 10) and KO8A (SEQ ID NO:
- 11).
A set of oligonucleotides primers for the co-amplification of nucleic acid from Treponema pallidum and Haemophilus ducreyi, wherein said set comprises:
- (a) a pair of primers for the polymerase chain reaction amplification of a region of the Treponema pallidum 47 kilodalton membrane immunogen gene, wherein said pair of primers is KO3A (SEQ ID NO:
- 6) and KO4 (SEQ ID NO:
- 7);
- and
- (b) a pair of primers for the polymerase chain reaction amplification of a region of the Haemophilus ducreyi 16S ribosomal RNA gene, wherein said pair of primers is KO7A (SEQ ID NO:
- 10) and KO8A (SEQ ID NO:
- 11).
An oligonucleotide probe for the detection and identification of nucleic acid from Treponema pallidum, wherein said probe hybridizes under sequence-specific hybridization conditions to a region of the Treponema pallidum 47 kilodalton membrane immunogen gene, wherein said probe is selected from the group consisting of KO 17 (SEQ ID NO:
- 8), KO18 (SEQ ID NO:
- 9), and the complements thereof.
An oligonucleotide probe for the detection and identification of nucleic acid from Haemophilus ducreyi, wherein said probe hybridizes under sequence-specific hybridization conditions to a region of the Haemophilus ducreyi 16S ribosomal RNA gene, wherein said probe is KO15 (SEQ ID NO:
- 12) or the complement thereof.
A set of oligonucleotides for the amplification, detection, and identification of nucleic acid from Treponema pallidum, wherein said set comprises:
- (a) a pair of primers for the polymerase chain reaction amplification of a region of the Treponema pallidum 47 kilodalton membrane immunogen gene, wherein said pair of primers is KO3A (SEQ ID NO:
- 6) and KO4 (SEQ ID NO:
- 7);
- (b) an oligonucleotide probe for the detection and identification of nucleic acid from Treponema pallidum, wherein said probe hybridizes under sequence-specific hybridization conditions to a region of the Treponema pallidum 47 kilodalton membrane immunogen gene, wherein said probe is selected from the group consisting of KO17 (SEQ ID NO:
- 8), KO18 (SEQ ID NO:
- 9), and the complements thereof.
A set of oligonucleotides for the amplification, detection, and identification of nucleic acid from Haemophilus ducreyi, wherein said set comprises:
- (a) a pair of primers for the polymerase chain reaction amplification of a region of the Haemophilus ducreyi 16S ribosomal RNA gene, wherein said pair of primers is KO7A (SEQ ID NO:
- 10) and KO8A (SEQ ID NO:
- 11);
- and
- (b) an oligonucleotide probe for the detection and identification of nucleic acid from Haemophilus ducreyi, wherein said probe hybridizes under sequence-specific hybridization conditions to a region of the Haemophilus ducreyi 16S ribosomal RNA gene, wherein said probe is KO15 (SEQ ID NO:
- 12) or the complement thereof.
A set of oligonucleotides for the co-amplification, detection, and identification of nucleic acid from Treponema pallidum and Haemophilus ducreyi, wherein said set comprises:
- (a) a pair of primers for the polymerase chain reaction amplification of a region of the Treponema pallidum 47 kilodalton membrane immunogen gene, wherein said pair of primers is KO3A (SEQ ID NO:
- 6) and KO4 (SEQ ID NO:
- 7);
- (b) an oligonucleotides probe for the detection and identification of nucleic acid from Treponema pallidum, wherein said probe hybridizes under sequence-specific hybridization conditions to a region of the Treponema pallidum 47 kilodalton membrane immunogen gene, wherein said probe KO17 (SEQ ID NO:
- 8) or KO18 (SEQ ID NO:
- 9);
- (c) a pair of primers for the polymerase chain reaction amplification of a region of the Haemophilus ducreyi 16S ribosomal RNA gene, wherein said pair of primers is KO7A (SEQ ID NO:
- 10) and KO8A (SEQ ID NO:
- 11);
- and
- (d) an oligonucleotides probe for the detection and identification of nucleic acid from Haemophilus ducreyi, wherein said probe hybridizes under sequence-specific hybridization conditions to a region of the Haemophilus ducreyi 16S ribosomal RNA gene, wherein said probe is KO15 (SEQ ID NO:
- 12).
Dependencies are inferred from the claim text.