COMPOSITIONS FOR DETECTING SMALL RNAS

摘要

Compositions and reaction mixtures are provided for the detection of small RNA target nucleic acids, preferably miRNA target nucleic acids, wherein the compositions and reaction mixtures provide for sensitive and specific detection of the target nucleic acids. The compositions and reaction mixtures include one or more of a first amplification oligomer that is preferably an extender primer, a target capture oligomer that is preferably at least partially double stranded, a promoter primer/provider, a reverse primer that is preferably a universal primer and a detection probe. The compositions and reaction mixtures are useful for diagnostics, prognostics, monitoring the effectiveness of treatment and/or determining a treatment.

主权项

1. An amplification reaction mixture comprising:
(a) a miR-221 target capture probe that forms under hybridizing conditions at least a partially double-stranded structure that is a hairpin structure made up of a contiguous linear sequence that includes an internal target hybridizing sequence that is substantially complementary to all or a portion of SEQ ID NO:44 and that forms a single-stranded loop portion of the hairpin structure, wherein the internal target hybridizing sequence is flanked by a capture region and by a terminal region that binds to the capture region to form a double-stranded stem portion of the hairpin structure, wherein the capture region further comprises a substantially homopolymeric nucleic acid sequence and the terminal region further comprises a substantially homopolymeric nucleic acid sequence that is complementary to the capture region sequence, and wherein the miR-221 target capture probe is at least 95% identical to SEQ ID NO:24; and (b) a first amplification oligomer configured to hybridize to a miR-221 target nucleic acid that is an extender primer comprising a 3′ target hybridizing sequence and a 5′ extension nucleic acid sequence, wherein the 3′ target hybridizing sequence is substantially identical to about 9 or more contiguous nucleic acids at the 5′ end of the target hybridizing sequence of the miR-221 target capture probe.