Appeal 2007-2363
Application 10/253,967
1. A method for detecting the presence of a target nucleic acid sequence
on a sample nucleic acid strand comprising the steps of:
contacting a sample suspected of containing said target nucleic acid
sequence with a diagnostic probe under hybridizing conditions;
wherein the nucleotide sequence of said diagnostic probe comprises
(1) a first probe region at its 5'-end that is substantially complementary to a
first target region characteristic of said target nucleic acid sequence, and (2)
a second probe region, located 3' to said first probe region, where the second
probe region is substantially complementary to a second target region
characteristic of said target nucleic acid sequence on the target nucleic acid
strand wherein the first and second probe regions on the diagnostic probe
may be separated by a spacer region of nucleic acid, and further there exists
an intervening sequence between the first and second target regions on the
target nucleic acid strand;
wherein when said first and second probe regions are separated by a
spacer region then said spacer region forms a non-self-hybridized loop under
said selected conditions;
whereby for said selected hybridization conditions the first and second
probe regions are such that the diagnostic probe is only stably hybridized to
the target nucleic acid strand to form a detectable probe:target hybrid when
the first probe region is substantially complementary to the first target region
and the second probe region is substantially complementary to the second
target region,
but wherein for said selected hybridization conditions the diagnostic
probe is not stably hybridized to the target nucleic acid strand to form a
probe:target hybrid detectable above a threshold indicative of stable
hybridization when either the first probe region is not substantially
complementary to the first target region or the second probe region is not
substantially complementary to the second target region; and
detecting the presence or absence of the stable probe:target hybrid in
the absence of elongation of the probe:target hybrid as an indication of the
presence of the target nucleic acid sequence in the sample.
9. A method for detecting the presence of two or more target nucleic
acid sequences on two or more sample nucleic acid strands comprising the
steps of:
contacting a sample suspected of containing said target nucleic acid
sequences with a diagnostic probe under hybridizing conditions;
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Last modified: September 9, 2013