Modified N-4 nucleotides for use in amplified nucleic acid hybridization assays

• US 5,594,118 A
• Filed: 05/10/1995
• Issued: 01/14/1997
• Est. Priority Date: 10/15/1987
• Status: Expired due to Term

• Current Assignee: Siemens Healthcare Diagnostics Incorporated (Siemens AG)
• Original Assignee: Chiron Corporation (Bayer AG)
• Inventors: Urdea, Michael S., Horn, Thomas
• Primary Examiner(s): Jones, W. Gary
• Assistant Examiner(s): REES, DIANE
• Application Number:
  US08/438,413
• Time in Patent Office: 615 Days
• Field of Search: 536/23.1, 536/24.3, 435/6, 435/5, 435/91.1
• US Class Current: 536/23.1
• CPC Class Codes: C07H 19/06 Pyrimidine radicals
• 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 1/682 Signal amplification
• CPC Class Codes: C12Q 1/689 for bacteria
• CPC Class Codes: C12Q 1/706 for hepatitis
• CPC Class Codes: G01N 2333/045 Cytomegalovirus
• CPC Class Codes: G01N 33/532 Production of labelled immunochemicals
• CPC Class Codes: Y10S 435/81 Packaged device or kit
• CPC Class Codes: Y10S 435/948 using viruses or cell lines
• CPC Class Codes: Y10S 436/811 Test for named disease, body condition or organ function

Abstract

Linear or branched oligonucleotide multimers useful as amplifiers in biochemical assays which comprise (1) at least one first single-stranded oligonucleotide unit that is complementary to a single-stranded oligonucleotide sequence of interest, and (2) a multiplicity of second single-stranded oligonucleotide units that are complementary to a single-stranded labeled oligonucleotide. Amplified sandwich nucleic acid hybridizations and immunoassays using the multimers are exemplified.

Claims

1. A nucleic acid having the structure ##STR12## where Z is a nucleophile, R¹ is a protective group that is base-stable and acid sensitive, R² is methyl, R³ is a protective group that can be removed and replaced with hydrogen without affecting R¹, R⁵ is a phosphorus derivative that enables addition of nucleotides to the 5'"'"' position of an oligonucleotide chain during chemical synthesis, R⁶ is methyl, hydrogen, I, Br or F, and X is an integer in the range of 1 to 8, inclusive.

2. The nucleic acid of claim 1, wherein Z is selected from the group consisting of --O--, --NH--, --S--, PO₄ ═

and --O(CO)O--.

3. The nucleic acid of claim 1, wherein R¹ is dimethoxytrityl or pixyl.

5. The nucleic acid of claim 1, wherein R⁵ is a phosphoramidite, a phosphodiester or a phosphotriester.

4. A nucleic acid having the structure ##STR13## where Z is a nucleophile, R¹ is a protective group that is generally base-stable and acid sensitive, R² is hydrogen or methyl, R³ is a protective group that can be removed and replaced with hydrogen without affecting R¹, R⁵ is a phosphorus derivative that enables addition of nucleotides to the 5'"'"' position of an oligonucleotide chain during chemical synthesis, R⁶ is methyl hydrogen I, Br or F, and X is an integer in the range of 1 to 8, inclusive, wherein R³ is selected from the group consisting of ##STR14##