Abbott Laboratories Corporation

Abbott Molecular Inc.

Thomas G Laffler

John J Carrino

Ronald L Marshall

Stanley R Bouma

Larry G Birkenmeyer

Jon D Kratochvil

Natalie A Solomon

Keith C Backman

Bruce J Dille

Hsiang-Yun Y Hu

Uwe Spies

Laurie A Rinehardt

Jeffrey L Joseph

Sheila B Bond

A method and kits for amplifying and detecting target nucleic acid sequences in a sample is disclosed. The method employs primers which have reactive pair members linked to them. The reactive pair members can be attached to a solid phase and/or detected by labeled conjugate.

Detection and amplification of target nucleic acid sequences

The present invention involves methods of improving LCR and PCR amplification schemes by modifying at least one probe/primer end so that the probability of the probe/prirner contributing to spurious signal development is greatly reduced. Only after specific hybridization of the modified probe/primer with true target, are the modified ends 'corrected' in a target dependent fashion to allow participation of the probe/primer in the enzymatic assembly reaction. Specific modifications depend on whether the assembly is done by ligation (as in LCR) or by extension/elongation (as in PCR).

Method and kits for amplifying target nucleic acids applicable to both

Short nucleotide sequences of human papilloma virus useful for the determination of the presence and type of human papilloma virus present in a test sample. The sequences provided can be amplified by polymerase chain reaction or ligase chain reaction. The sequences provided also can be hybridized by standard slot-, dot- or replica-blot procedures. Methods and kits also are provided for the detection of human papilloma virus in a test sample and the determination of the type of human papilloma virus present in the test sample.

Nucleotide sequences useful as type specific probes, PCR primers and LCR

An improved, 'gap filling' embodiment of the Ligase Chain Reaction (LCR) is described. Gap filling LCR is LCR wherein at least one of the probes is recessed so that a gap is formed between the adjacent probes when they are hybridized to target. The gap is filled using polymerase and deoxyribonucleotide triphosphates before ligation of the probes together. There are single and double gap versions, depending on whether one or two probes are recessed and require filling before ligation. The improvement resides in selecting and using target sequences such that only a single type, or two types, of deoxyribonucleotide triphosphate(s) are required to fill double gaps each being 1-10 bases in length, preferably 1-3 bases. Probes having specific sequences are claimed for a number of pathogens.

Amplification of target nucleic acids using gap filling ligase chain

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Libertyville, IL, United States of America

Thomas G Laffler