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1. WO2000014281 - ASSAYS USING CROSSLINKABLE IMMOBILIZED NUCLEIC ACIDS

Publication Number WO/2000/014281
Publication Date 16.03.2000
International Application No. PCT/US1999/018950
International Filing Date 23.08.1999
Chapter 2 Demand Filed 09.03.2000
IPC
C12Q 1/68 2006.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
1Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
68involving nucleic acids
CPC
C12Q 1/6827
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
1Measuring or testing processes involving enzymes, nucleic acids or microorganisms
68involving nucleic acids
6813Hybridisation assays
6827for detection of mutation or polymorphism
C12Q 1/6832
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
1Measuring or testing processes involving enzymes, nucleic acids or microorganisms
68involving nucleic acids
6813Hybridisation assays
6832Enhancement of hybridisation reaction
C12Q 1/6837
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
1Measuring or testing processes involving enzymes, nucleic acids or microorganisms
68involving nucleic acids
6813Hybridisation assays
6834Enzymatic or biochemical coupling of nucleic acids to a solid phase
6837using probe arrays or probe chips
C12Q 1/6841
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
1Measuring or testing processes involving enzymes, nucleic acids or microorganisms
68involving nucleic acids
6813Hybridisation assays
6841In situ hybridisation
Applicants
  • NAXCOR [US]/[US]
Inventors
  • HUAN, Bingfang
  • ALBAGLI, David
  • WOOD, Michael, L.
  • VAN ATTA, Reuel, B.
  • CHENG, Peter, C.
Agents
  • CHOW, Y., Ping
Priority Data
09/138,19521.08.1998US
Publication Language English (EN)
Filing Language English (EN)
Designated States
Title
(EN) ASSAYS USING CROSSLINKABLE IMMOBILIZED NUCLEIC ACIDS
(FR) DOSAGES UTILISANT DES ACIDES NUCLEIQUES RETICULABLES IMMOBILISES
Abstract
(EN)
Improved methods for $i(in situ) hybridization assays of cellular and subcellular systems and tissue sections, and immobilization-based assay techniques such as Northern blotting, Southern blotting, dot blots, and the like, and assay techniques wherein the probes are bound to substrates are disclosed. The subject invention employs crosslinker containing hybridization probes capable of forming covalent bonds between the probe and the target nucleic acid. Upon activation, the crosslinker will, if the probe has hybridized with its essentially complementary target, form covalent bonds with the complementary strand to covalently crosslink the probe to the target. Subsequently, stringent wash conditions may be employed to reduce background signals due to non-specific absorption or probes or targets, while retaining all crosslinked probe/target hybrids. Also disclosed are diagnostic kits for use in clininical and diagnostic laboratories.
(FR)
L'invention concerne des procédés améliorés destinés aux dosages d'hybridation in situ de systèmes cellulaires et sous-cellulaires et de parties de tissus, ainsi que des techniques de dosage fondées sur l'immobilisation telles que Northern blot, Southern blot et dot blot, de même que des techniques de dosage dans lesquelles les sondes sont liées aux substrats. L'invention utilise des sondes d'hybridation employant des agents de réticulation capables de former des liaisons covalentes entre la sonde et l'acide nucléique cible. Après activation, si la sonde s'est hybridée avec sa cible sensiblement complémentaire, l'agent de réticulation va former des liaisons covalentes avec le brin complémentaire pour réticuler de manière covalente la sonde et la cible. Par la suite, on peut recourir à des conditions strictes de lavage pour réduire les signaux d'arrière-plan dus à l'absorption non spécifique ou aux sondes ou encore aux cibles, et ce tout en préservant tous les hybrides réticulés sonde/cible. L'invention concerne aussi des kits diagnostiques utilisés dans des laboratoires cliniques et diagnostiques. -
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