Compositions and methods to detect Enterococci nucleic acid

• US 7,588,897 B2
• Filed: 05/11/2007
• Issued: 09/15/2009
• Est. Priority Date: 05/12/2006
• Status: Active Grant

• Current Assignee: Gen-Probe Incorporated (Hologic Incorporated)
• Original Assignee: Gen-Probe Incorporated (Hologic Incorporated)
• Inventors: Livezey, Kristin W., Pollner, Reinhold B.
• Primary Examiner(s): Johannsen; Diana B
• Application Number:
  US11/747,314
• Publication Number:
  US 20070264658A1
• Time in Patent Office: 858 Days
• Field of Search: None
• US Class Current: 435/6.15
• CPC Class Codes: C12Q 1/689 for bacteria

Abstract

The disclosed invention includes nucleic acid oligomers that may be used as amplification oligomers, including primers, capture probes for sample preparation, and detection probes for detection of indicator enterococci 23S rRNA sequences in samples by using methods of specific nucleic acid amplification and detection.

Claims

1. A method for amplifying and detecting indicator enterococci nucleic acid in a sample, wherein said indicator enterococci nucleic acid is nucleic acid from E. faecalis, E. faecium, E. casseliflavus, E. gallinarum, E. mundtii, E. durans, E. hirae and/or E. columbae and not from E. avium, E. malodoratus, E. pseudoavium, E. raffinosus, E. saccharolyticus or E. dispar, comprising the steps of:

a) contacting said sample with a first amplification oligomer comprising a target binding region consisting of SEQ ID NO;

30, a second amplification oligomer comprising a target binding region consisting of SEQ ID NO;

31, and a detection probe oligomer comprising a target binding region consisting of SEQ ID NO;

45;

b) exposing said sample to conditions sufficient to amplify indicator enterococci nucleic acid if present in said sample with said first and second amplification oligomers to produce an amplified product; and

c) determining whether said indicator enterococci nucleic acid is in said sample by using said detection probe oligomer to detect the presence or absence of the amplified product.

2. The method of claim 1, wherein said detection probe oligomer comprises at least one non-target binding region.

3. The method of claim 2, wherein said detection probe oligomer comprises two non-target binding regions tat hybridize to each other when said detection probe oligomer is not hybridized to indicator enterococci nucleic acid.

4. The method of claim 1, 2 or 3, wherein said detection probe oligomer comprises a detectable label.

5. The method of claim 4, wherein said detection probe oligomer comprises a fluprophore and a quencher.

6. The method of claim 1, wherein said second amplification oligomer comprises a promoter sequence.

7. The method of claim 6, wherein said promoter sequence consists of SEQ ID NO:

49.

17. The method of claim 1, wherein the sample is an enviromnental sample or a biological sample.

18. The method of claim 1, wherein the sample is a processed sample.

8. A kit for amplifying and detecting indicator enterococci nucleic acid in a sample, wherein said indicator enterococci nucleic acid is nucleic acid from E. faecalis, E. faecium, E. casseliflavus, E. gallinarum, E. mundtii, E. durans, E. hirae and/or E. columbae and not from E. avium, E. malodoratus, E. pseudoavium, E. raffinosus, E. saccharolvticus or E. disbar, comprising:

a first amplification oligomer 27-100 nucleotides in length comprising a target binding region consisting of SEQ ID NO;

30;

a second amplification oligomer 34-100 nucleotides in length comprising a target binding region consisting of SEQ ID NO;

31; and

a detection probe oligomer 15-100 nucleotides in length comprising a target binding region consisting of SEQ ID NO;

45.

9. The kit of claim 8, wherein said detection probe oligomer 15-100 nucleotides in length comprises at least one non-target binding region.

10. The kit of claim 9, wherein said detection probe oligomer 15-100 nucleotides in length comprises two non-target binding regions that hybridize to each other when said detection probe oligomer is not hybridized to indicator enterococci nucleic acid.

11. The kit of claim 8, 9 or 10, wherein said detection probe oligomer 15-100 nucleotides in length comprises a detectable label.

12. The kit of claim 11, wherein said detection probe oligomer 15-100 nucleotides in length comprises a fluorophore and a quencher.

13. The kit of claim 8, wherein said second amplification oligomer 34-100 nucleotides in length comprises a promoter sequence.

14. The kit of claim 13, wherein said promoter sequence consists of SEQ ID NO:

49.

15. The kit of claim 8, further comprising a capture oligomer.

16. The kit of claim 15, wherein said capture oligomer comprises SEQ ID NO:

50, 51 or 52.