| US 7,601,497 B2 | ||
| Detection of nucleic acids by target-specific hybrid capture method | ||
| Irina Nazarenko, Gaithersburg, Md. (US); Attila Lorincz, North Potomac, Md. (US); Paul Eder, Bethesda, Md. (US); Brian Lowe, Olney, Md. (US); Richard Mallonee, Baltimore, Md. (US); and Ha Thai, Chevy Chase, Md. (US) | ||
| Assigned to Qiagen Gaithersburg, Inc., Gaithersburg, Md. (US) | ||
| Filed on Nov. 07, 2005, as Appl. No. 11/269,003. | ||
| Application 11/269003 is a continuation in part of application No. 11/005617, filed on Dec. 06, 2004, abandoned. | ||
| Application 11/005617 is a continuation in part of application No. 10/971251, filed on Oct. 20, 2004, abandoned. | ||
| Prior Publication US 2006/0051809 A1, Mar. 09, 2006 | ||
| Int. Cl. C12Q 1/68 (2006.01); C12P 19/34 (2006.01); C07H 21/04 (2006.01) | ||
| U.S. Cl. 435—6 [435/91.2; 536/24.31; 536/24] | 26 Claims |
| 1. A method of detecting at least one target nucleic acid comprising:
a) forming a plurality of enriched target nucleic acids comprising
i) mixing the at least one target nucleic acid with a nucleic acid probe complementary to at least a portion of the at least
one target nucleic acid to form a target DNA:RNA hybrid, wherein when the target nucleic acid is RNA, the probe is DNA, and
wherein when the target nucleic acid is DNA, the probe is RNA;
ii) capturing the target DNA:RNA hybrid onto a solid support; and
iii) removing any nucleic acid that does not form the target DNA:RNA hybrid or that is not captured onto the solid support
to form the plurality of enriched target nucleic acids;
b) amplifying the plurality of enriched target nucleic acids from step a) to form a plurality of amplified targets; and
c) detecting the at least one target nucleic acid in the plurality of amplified targets by
i) mixing the plurality of amplified targets with a selectable oligonucleotide and a nucleic acid probe, wherein the selectable
oligonucleotide hybridizes to a first portion of the amplified targets and wherein the nucleic acid probe is complementary
to a second portion of the amplified targets wherein if the amplified target is DNA, the selectable oligonucleotide and the
nucleic acid probe is RNA and if the amplified target is RNA, the selectable oligonucleotide and the nucleic acid probe is
DNA to form a DNA:RNA hybrid target/oligonucleotide/probe complex, wherein the first portion of the amplified targets and
the second portion of the amplified targets are different; and
ii) detecting the DNA:RNA hybrid target/oligonucleotide/probe complex by a DNA:RNA hybrid-specific binding agent, to detect
at least one target nucleic acid.
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