Biomerieux, Inc.

Francedillaois Mallet

Bernard F Mandrand

Philippe L Cleuziat

Claude Mabilat

Philippe Cros

Guy Oriol

Pierre Levasseur

William McAllister

Patrice A Allibert

Francedillaoise Guillou-Bonnici

A chimera oligonucleotide is provided that can be used in a process for obtaining transcripts and/or amplification of a target sequence of a nucleic acid, having, at its 3' end, a downstream sequence. The oligonucleotide comprises successively, from 5' to 3', 1) a first oligonucleotide segment, of the DNA type, comprising a sense sequence of a promoter of an RNA polymerase, 2) a second oligonucleotide segment, of the DNA type, capable of hybridizing with the downstream sequence, and 3) a third oligonucleotide segment, of the RNA type, capable of hybridizing with a part of the target sequence contiguous to the downstream sequence, the third segment being blocked at 3'. A process using the chimera oligonucleotide and an enzyme system containing DNA polymerase activity, RNA polymerase activity, and a third activity, for example, an RNase H activity provides transcription products of the target. By adding a second chimera oligonucleotide capable of hybridizing with the complement of the target, cyclic amplification of the target and its complement are obtained.

Chimera oligonucleotide and its utilization for obtaining transcripts of

A procedure for detecting single-stranded nucleotide sequences in a sample is disclosed wherein a passively fixed capture probe is used in concert with a non-radioactively labelled detection probe in a sandwich hybridization technique.

Sandwich hybridization assays using very short capture probes

An RNA amplification method is particularly useful for diagnosing bacterial or viral infections or genetic disorders and for cell typing. The method includes the steps of denaturing a solution containing RNA, synthesizing a first cDNA strand from a suitable primer in the presence of reverse transcriptase, denaturing the heteroduplex formed, synthesizing a second cDNA strand from a second primer in the presence of DNA polymerase and then subjecting the cDNA formed to a sufficient number of amplification cycles. All the reactants and solvents are first placed in the same container to provide a single manipulation step that avoids the risk of contamination.

RNA amplification method requiring only one manipulation step

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Villeurbanne, France

Francedillaois Mallet