Method for detection of specific nucleic acid sequences

• US 5,516,641 A
• Filed: 03/10/1995
• Issued: 05/14/1996
• Est. Priority Date: 08/25/1988
• Status: Expired due to Term

• Current Assignee: Siemens Healthcare Diagnostics Products GmbH (Siemens AG)
• Original Assignee: Syntec Incorporated (Atos SE)
• Inventors: Stull, Paul D., Ullman, Edwin F., Goodman, Thomas C.
• Primary Examiner(s): Zitomer, Stephanie W.
• Application Number:
  US08/401,660
• Time in Patent Office: 431 Days
• Field of Search: 435/6, 935/77, 935/78
• US Class Current: 435/6.12
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• CPC Class Codes: C12Q 1/6816 characterised by the detection means C12Q1/6804 takes precedence
• 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 2523/101 Crosslinking agents, e.g. psoralen
• CPC Class Codes: C12Q 2533/101 Primer extension
• CPC Class Codes: C12Q 2533/107 Probe or oligonucleotide ligation
• CPC Class Codes: C12Q 2565/101 Interaction between at least two labels
• CPC Class Codes: C12Q 2565/501 being an array of oligonucleotides
• CPC Class Codes: Y10S 435/81 Packaged device or kit
• CPC Class Codes: Y10S 435/975 Kit
• CPC Class Codes: Y10T 436/143333 Saccharide [e.g., DNA, etc.]

Abstract

A kit is disclosed for a method for detecting the presence of a target polynucleotide sequence. The kit comprises a first polynucleotide sequence and a second polynucleotide sequence complementary to non-contiguous portions of a target polynucleotide sequence, which first and second sequences are covalently attached when they are hybridized to the target sequence. The presence of the covalently attached first and second sequences is related to the presence of the target polynucleotide sequence. The invention may be applied to target polynucleotide sequences in DNA or RNA. Specific target polynucleotide sequences of interest will frequently be characteristic of particular microorganisms, viruses, viroids, or genetic characteristics, including genetic abnormalities.

Claims

1. A kit for use in detecting a target polynucleotide sequence, which comprises in packaged combination:

a first polynucleotide complementary to and capable of hybridizing to a portion of a single strand of a target polynucleotide and having, or capable of having, means for immobilizing said first polynucleotide,

a second polynucleotide, having, or capable of having, a label wherein said second polynucleotide is complementary to and capable of hybridizing to a portion of said single strand of said target polynucleotide other than and non-contiguous with the portion complementary to said first polynucleotide, and

means for covalently attaching said first and second polynucleotides when said first and second polynucleotides are hybridized with said single strand of said target polynucleotide.

2. The kit of claim 1 wherein one of said first and said second polynucleotides is capable of undergoing an extension reaction in the presence of a polynucleotide polymerase and one or more deoxynucleoside triphosphates to become contiguous with the other of said first and said second polynucleotides when both first and second polynucleotides are hybridized to said single strand of said target polynucleotide, and said kit further comprises:

a polynucleotide polymerase, one or more deoxynucleoside triphosphates, and a ligase as a means for covalently attaching said first and second polynucleotides.