| 摘要 |
Reagents, systems and methods for the detection of nucleic acids are described herein, including such methods which may be performed in a single reaction. Such reagents include a target specific indexing probe (TSIP) synthetic DNA structure comprising a detectable moiety and a quencher thereof, wherein in the presence of a target nucleic acid a portion (a tag) of the (TSIP) synthetic DNA structure comprising the detectable moiety is released, thereby increasing the signal emitted therefrom. The tag may in turn bind to a capture probe, facilitating signal detection. |
| 主权项 |
1. A method for detecting the presence of a target nucleic acid in a sample, the method comprising:
(a) contacting a sample with amplification and hybridization reagents, wherein said amplification and hybridization reagents comprise a target specific indexing probe (TSIP) synthetic DNA structure comprising a first segment, a second segment and a third segment, wherein:
(i) the first segment comprises a sequence complementary to a first portion of the target nucleic acid and complementary to the third segment;(ii) the second segment is attached to the 3′ end of the first segment and comprises a sequence that is complementary to a second portion of the target nucleic acid that is contiguous to the first portion, but not complementary to the first or third segment;(iii) the third segment is attached to the 3′ end of the second segment and comprisesa first part comprising a sequence that is complementary to the first segment and to a capture probe present in said reaction;a first part comprising a sequence that is complementary to the first segment, and a second part comprising a sequence that is complementary to the capture probe;ora first part comprising a sequence that is complementary to the first segment and to a first portion of the capture probe, and a second part comprising a sequence that is complementary to a second portion of the capture probe, wherein the third segment comprises a detectable label attached thereto, and said first and/or second segment comprises a quenching moiety for the detectable label, wherein the detectable label and the quenching moiety are located within sufficiently close proximity of each other in the TSIP synthetic DNA structure such that the signal emitted by the detectable label is reduced as compared to a corresponding signal emitted by a detectable label in a corresponding TSIP synthetic DNA structure lacking said quencher moiety; (b) performing an amplification and hybridization reaction, wherein said third segment is released from the first and second segment if the target nucleic acid is present; and (c) detecting the hybridization of the third segment to the capture probe by virtue of an increase in the signal emitted from the detectable moiety relative to the signal emitted from a corresponding TSIP that has not been contacted with the sample, wherein the detection of an hybridization is indicative that the target nucleic acid is present in the sample. |
