| 摘要 |
Methods and compositions are provided for assaying for a target analyte in a sample by formation of a substrate for a replicase. The target analyte, if present in the sample, is first bound to a reporter probe. The reporter probe comprises a first portion and a second portion. The first portion comprises a polynucleotide that encodes at least part of a sequence for an RNA that is a substrate for replication. The second portion comprises a molecule that has affinity for an analyte. The reporter probe itself is not a substrate for a replicase. However, a replicase substrate is generated by treating reporter probe which is bound to analyte with a composition having nuclease activity in order to release the parts of the first portion, and then the released parts and one or more mononucleotides or oligonucleotides comprising missing parts of the substrate sequence are joined with a composition having ligase activity. Newly formed substrate is replicated with a replicase, and a signal is measured which indicates the presence and quantity of analyte in the sample. The invention also provides methods and compositions for simultaneously assaying for any of multiple analytes in a sample.
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