Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Patent
1993-02-05
1995-04-18
Zitomer, Stephanie W.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
435 911, 935 77, 935 78, C12Q 168, C12P 1934
Patent
active
054077982
ABSTRACT:
New methods are provided for the amplification of a midivariant DNA containing an inserted target specific nucleic acid sequence(s) to enable detection of the presence of a target nucleic acid sequence in a test sample. One method employs midivariant DNA as a template for the synthesis of RNA catalyzed by QB replicase. Two midivariant DNA/probe conjugates including a nonreplicable portion of midivariant DNA and a target specific nucleic acid sequence (probe) are described. An amplification method including the steps of hybridization and ligation of the midivariant DNA/probe conjugates followed by replication of the DNA template has enabled detection of less than 200 template molecules. In a modified amplification method one of the midivariant DNA/probe conjugates further includes a RNA polymerase promoter sequence to enable transcription of the midivariant DNA template into RNA. The sequential ligation-transcription-replication method enables the detection of a single template molecule. The detectable molecule(s) are indicative of the presence of the target nucleic acid sequences in the test sample.
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Donahue Jeffrey J.
Martinelli Richard A.
Unger John T.
Ciba Corning Diagnostics Corp.
Morgenstern Arthur S.
Roesler Judith A.
Slepchuk, Jr. Nicholas I.
Zitomer Stephanie W.
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