Intergenic spacer target sequence for detecting and distinguishing Chlamydial species or strains
AssigneeThe United States of America as represented by the Secretary of Agriculture
InventorsKarin D. E. Everett, Arthur A. Andersen
Life of this patent
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Abstract
The intergenic spacer between the 16S and the 23S rRNA genes and a 131-bp segment at the 3′ end of Domain I of the 23S gene provides suitable target sequences for developing probes and primers useful in detecting and identifying Chlamydiaceae in laboratory and clinical samples.
Claims12
An isolated oligonucleotide useful as a primer wherein the oligonucleotide consists of a sequence selected from the group consisting of SEQ ID NO:
- 55, SEQ ID NO:
- 61 and SEQ ID NO:
- 67, the complement of SEQ ID NO:
- 55, SEQ ID NO:
- 61 and SEQ ID NO:
- 67, and the RNA equivalent of SEQ ID NO:
- 55, SEQ ID NO:
- 61 and SEQ ID NO:
- 67.
An isolated oligonucleotide from 15-100 nucleotides in length comprising a sequence selected from the group consisting of SEQ ID NOs. 56 and 59-60, 62-66, and 68-73, the complement of each of said sequences, and the RNA equivalent of each of said sequences.
A method for distinguishing in a sample a first species or strain of Chlamydiaceae set forth in Table IV from all other species or strains of Chlamydiaceae set forth in Table IV, said method comprising:
- a. amplifying nucleic acid contained in said sample with at least one species-specific or strain-specific primer having a length of about 15-30 nucleotides that hybridizes under high stringency conditions to a nucleic acid segment selected from the group consisting of Region A of chlamydial rDNA as defined in Table IV, Region B of chlamydial rDNA as defined in Table IV, the complement of said Region A, the complement of said Region B, the RNA equivalent of said Region A, and the RNA equivalent of said Region B, and thereby producing an amplified nucleic acid segment wherein said segment is specific to said first species or strain;
- and
- b. detecting the presence of said amplified segment as indicative of said first species or strain of Chlamydiaceae distinguished from all other species or strains of Chlamydiaceae.
A method for distinguishing in a sample a first species or strain of Chlamydiaceae set forth in Table IV from all other species or strains of Chlamydiaceae set forth in Table IV, said method comprising:
- a. contacting said sample with a labeled, single-stranded species-specific or strain-specific probe having a length of at least about 15 nucleotides and up to about 100 nucleotides that will hybridize with a nucleic acid segment selected from the group consisting of Region A of chlamydial rDNA as defined in Table IV, Region B of chlamydial rDNA as defined in Table IV, the complement of said Region A, the complement of said Region B, the RNA equivalent of said Region A, and the RNA equivalent of said Region B, and thereby producing a species-specific or strain-specific hybridization product;
- and
- b. detecting said hybridization product as indicative of said first species or strain of Chlamydiaceae distinguished from all other species or strains of Chlamydiaceae.
Dependencies are inferred from the claim text.