| US 7,468,261 B2 | ||
| Molecular detection systems utilizing reiterative oligonucleotide synthesis | ||
| Michelle M Hanna, Phoenix, Ariz. (US) | ||
| Assigned to Ribomed Biotechnologies, Inc., Phoenix, Ariz. (US) | ||
| Filed on Jun. 24, 2003, as Appl. No. 10/602,045. | ||
| Application 10/602045 is a division of application No. 09/984664, filed on Oct. 30, 2001, granted, now 7,045,319, filed on May 16, 2006. | ||
| Prior Publication US 2004/0234996 A1, Nov. 25, 2004 | ||
| This patent is subject to a terminal disclaimer. | ||
| Int. Cl. C12P 19/34 (2006.01); C12Q 1/68 (2006.01) | ||
| U.S. Cl. 435—91.1 [435/6] | 52 Claims |
| 1. A method for detecting a target DNA or RNA polynucleotide, said method comprising:
(a) incubating a single-stranded target polynucleotide with a second polynucleotide comprising a sequence that hybridizes
to the single-stranded target polynucleotide forming a bubble complex, wherein said bubble complex comprises a first double-stranded
upstream region, a non-complementary single-stranded middle region, and a second double-stranded downstream region, an initiator,
an RNA polymerase, and a terminator, wherein said initiator hybridizes with said target polynucleotide;
(b) extending said initiator until said terminator is incorporated into an oligonucleotide transcript, by the process of abortive
transcription;
(c) repeating steps (a) and (b), thereby synthesizing multiple reiterative oligonucleotide transcripts from said target polynucleotide;
and
(d) detecting or quantifying said reiterative oligonucleotide transcripts, wherein the presence of said reiterative oligonucleotide
transcripts is indicative of the presence of said target DNA or RNA polynucleotide.
|