Primers and probes for detecting human papillomavirus and human beta globin sequences in test samples

• US 9,090,948 B2
• Filed: 09/30/2008
• Issued: 07/28/2015
• Est. Priority Date: 09/30/2008
• Status: Active Grant

• Current Assignee: Abbott Molecular Incorporated (Abbott Laboratories Incorporated)
• Original Assignee: Abbott Molecular Incorporated (Abbott Laboratories Incorporated)
• Inventors: Abravaya, Klara, Erickson, Brian J., Huang, Shihai X., Mak, Wai-Bing X., Salituro, John A., Tang, Ning
• Primary Examiner(s): Pande, Suchira
• Application Number:
  US12/241,119
• Publication Number:
  US 20100081124A1
• Time in Patent Office: 2,492 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/6886 for cancer immunoassay for cancer G01N33/574
• CPC Class Codes: C12Q 1/708 for papilloma
• CPC Class Codes: C12Q 2600/112 Disease subtyping, staging or classification
• CPC Class Codes: C12Q 2600/16 Primer sets for multiplex assays

Abstract

The present invention relates to primers, probes, primer sets, primer and probe sets, methods and kits for detecting human papillomaviruses, human beta globin sequences and human papillomaviruses and human beta globin sequences in a test sample.

Claims

1. A set of polynucleotides for detecting HPV types 16, 18, 31, 35, 39, 45, 51, 52, 58, 59, and 66 in a test sample, wherein the polynucleotides comprise:

(a) three different isolated forward primers and two different isolated reverse primers,

(i) wherein each isolated forward primer consists of a sequence selected from the group consisting of;

SEQ ID NO;

1, SEQ ID NO;

2, SEQ ID NO;

3, and full-length complements thereof;

(ii) wherein each isolated reverse primer consists of a sequence selected from the group consisting of;

SEQ ID NO;

4 and SEQ ID NO;

5, and full-length complements thereof; and

(b) eleven different isolated probes, wherein each probe consists of a sequence selected from the group consisting of;

SEQ ID NO;

8, SEQ ID NO;

9, SEQ ID NO;

10, SEQ ID NO;

12, SEQ ID NO;

13, SEQ ID NO;

14, SEQ ID NO;

15, SEQ ID NO;

16, SEQ ID NO;

18, SEQ ID NO;

19, SEQ ID NO;

20, and full-length complements thereof, wherein each probe is labeled with a detectable label;

wherein the detectable label comprises a fluorescent moiety attached at the 5′

end of the probe.

4. The set of polynucleotides of claim 1, wherein each probe further comprises a quencher moiety attached at its 3′

end.

2. A primer and probe kit for detecting HPV types 16, 18, 31, 35, 39, 45, 51, 52, 58, 59, and 66 in a test sample, wherein said kit comprises:

(a) three different isolated forward primers and two different isolated reverse primers,

(i) wherein each isolated forward primer consists of a sequence selected from the group consisting of;

SEQ ID NO;

1, SEQ ID NO;

2 and SEQ ID NO;

3 and full-length complements thereof;

(ii) wherein each isolated reverse primer consists of a sequence selected from the group consisting of;

SEQ ID NO;

4, SEQ ID NO;

5 and full-length complements thereof;

(b) eleven different isolated probes, wherein each isolated probe consists of a sequence selected from the group consisting of;

SEQ ID NO;

8, SEQ ID NO;

9, SEQ ID NO;

10, SEQ ID NO;

12, SEQ ID NO;

13, SEQ ID NO;

14, SEQ ID NO;

15, SEQ ID NO;

16, SEQ ID NO;

18, SEQ ID NO;

19, SEQ ID NO;

20, and full-length complements thereof, wherein each probe is labeled with a detectable label;

wherein the detectable label comprises a fluorescent moiety attached at the 5′

end of the probe;

(c) amplification reagents.

3. The primer and probe kit of claim 2, wherein the kit further comprises an isolated forward primer consisting of SEQ ID NO:

6 or a complement thereof, wherein the complement is identical in length to SEQ ID NO;

6, an isolated reverse primer consisting of SEQ ID NO;

7 or a complement thereof, wherein the complement is identical in length to SEQ ID NO;

7, and an isolated probe consisting of SEQ ID NO;

22 or a complement thereof, wherein the complement is identical in length to SEQ ID NO;

22, for detecting human beta globin in a test sample.

5. The primer and probe kit of claim 2, wherein each probe further comprises a quencher moiety attached at its 3′

end.