Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Patent
1995-01-24
1999-11-23
Brusca, John S.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
536 243, 536 2532, C12Q 168, C07H 2100
Patent
active
059898097
ABSTRACT:
The present invention provides a method for detecting the presence of a target polynucleotide in a sample. The method comprises the steps of (a) contacting the sample under hybridizing conditions with (i) a single-stranded polynucleotide probe capable of hybridizing to the target polynucleotide and comprising a polynucleotide and at least one intercalating molecule attached to a nucleotide of the polynucleotide by means of a linker arm, and (ii) a background-reducing reagent which chemically modifies the intercalating molecule when the probe to which it is attached is single-stranded; and (b) detecting a property change resulting from the intercalation of the intercalating molecule into a target-probe hybrid, thereby detecting the target polynucleotide. The intercalating molecule which is part of the polynucleotide probe induces a change in a property, in either the probe, the target polynucleotide or a target-probe hybrid. The property change can be detected, for example, by means of a generated signal which can be identified or quantified. The present invention can be employed in a heterogeneous (two step or two phase) assay using a support to immobilize the target or probe, and a washing step, and in a homogeneous (one step or one phase) assay using a hybridization solution. Also provided are a composition and a nucleic acid hybridization kit useful for detecting the presence of a target polynucleotide in a sample.
REFERENCES:
Bhavsar, et al., "Synthesis of Acridine Derivatives", 107 Chemical Abstracts, 667 (Abstract No. 96576U) 1987.
Arnold et al. Clin. Chem. 35 1588-1594 (1989) Assay Formats Involving Acridinium-Ester-Labeled DNA Probes.
Morrison et al. "Organic Chemistry" Allyn and Bacon, Inc. Boston (1973) pp. 765-775.
Brusca John S.
Enzo Diagnostics, Inc.
Fedus, Esq. Ronald C.
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