Detection of nucleic acids by target-specific hybrid capture method

• US 8,901,287 B2
• Filed: 10/09/2009
• Issued: 12/02/2014
• Est. Priority Date: 10/20/2004
• Status: Expired due to Fees

• Current Assignee: QIAGEN Gaithersburg, Inc. (Qiagen NV)
• Original Assignee: QIAGEN Gaithersburg, Inc. (Qiagen NV)
• Inventors: Nazarenko, Irina, Lorincz, Attila, Eder, Paul, Lowe, Brian, Mallonee, Richard, Thai, Ha
• Primary Examiner(s): Chunduru, Prabha
• Application Number:
  US12/576,425
• Publication Number:
  US 20100036104A1
• Time in Patent Office: 1,880 Days
• Field of Search: 536/24.31, 536/24.32, 435/6
• US Class Current: 536/24.32
• CPC Class Codes: C12Q 1/6804 Nucleic acid analysis using immunogens immunoassay G01N33/53
• CPC Class Codes: C12Q 1/6834 Enzymatic or biochemical coupling of nucleic acids to a solid phase
• CPC Class Codes: C12Q 1/6876 Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
• CPC Class Codes: C12Q 1/708 for papilloma
• CPC Class Codes: C12Q 2522/101 Single or double stranded nucleic acid binding proteins
• CPC Class Codes: C12Q 2525/121 incorporating both deoxyribonucleotides and ribonucleotides
• CPC Class Codes: C12Q 2525/186 incorporating a non-extendable or blocking moiety
• CPC Class Codes: C12Q 2537/119 Triple helix formation
• CPC Class Codes: C12Q 2537/143 Multiplexing, i.e. use of multiple primers or probes in a single reaction, usually for simultaneously analyse of multiple analysis
• CPC Class Codes: C12Q 2565/519 characterised by the capture moiety being a single stranded oligonucleotide
• CPC Class Codes: C12Q 2565/543 characterised by the use of two or more capture oligonucleotide primers in concert, e.g. bridge amplification

Abstract

Target-specific hybrid capture (TSHC) provides a nucleic acid detection method that is not only rapid and sensitive, but is also highly specific and capable of discriminating highly homologous nucleic acid target sequences. The method produces DNA:RNA hybrids which can be detected by a variety of methods.

Claims

1. A composition comprising:

(a) a plurality of at least one probe selected from the group consisting of SEQ ID NO;

9 to 21 and a full-length complement thereof;

(b) an HPV nucleic acid hybridized to at least a portion of the plurality of probe (a) to form an RNA;

DNA hybrid;

(c) at least one nucleic acid primer hybridized to the HPV nucleic acid;

(d) an antibody bound to the RNA;

DNA hybrid, wherein said antibody is bound to a solid support; and

(e) a blocker probe hybridized to at least a portion of the plurality of probes from (a).

2. The composition of claim 1 further comprising at least a second nucleic acid probe selected from the group consisting of SEQ ID NO:

1 to 8, SEQ ID NO 22 to 64, and a complement thereof.

3. The composition of claim 2 wherein probe (a) and the second nucleic acid probe are capture sequence probes specific for an HPV nucleic acid.

4. The composition of claim 3 further comprising at least one nucleic acid having a sequence that is complementary to at least a portion of probe (a) or the second nucleic acid probe or both.

5. The composition of claim 3 further comprising at least one signal sequence probe.

6. A bead set formed from a composition of claim 3.

7. A bead set formed from a composition of claim 1.

8. A bead set formed from a composition of claim 2.

9. The composition of claim 1 wherein said solid support is a bead.

10. The composition of claim 1 further comprising a DNA polymerase associated with the target nucleic acid.

11. The composition of claim 10, wherein said composition is in an environment suitable for performing an isothermal amplification.