| 摘要 |
<p>Method for detecting biomolecules that includes generation of labeled nucleic acid replicons. Method for detecting biomolecules (I) comprises: (a) coupling (I) to a substance (II) that is part of a nucleic acid replication apparatus; (b) binding the (I)-(II) complex (C) formed to immobilized binding molecules (III) that are specific for (I); (c) optionally washing to remove any unbound (C); (d) incubating bound (C) with: (i) high molecular weight nucleic acid (IV) and mononucleotides (MN), at least one of which is detectably labeled; and (ii) a second substance (V) that with the coupled (II) completes a functional replication apparatus for nucleic acid molecules (IVa) that bind to (IV), generating, by incorporation of labeled MN, replicates of (IV) that do not dissociate; (e) optionally removing (IV) and MN present in solution, by washing; and (f) determining (I) by detecting labeled replicates. Independent claims are also included for the following: (1) similar method in which immobilized (I) are incubated with a complex (C1) of (III) and (II), any unbound C1 is optionally removed by washing, then the (I)-C1 product formed is reacted as in steps (d)-(f) above; (2) method for preparing a marker for determining (I); and (3) markers prepared by method (2).</p> |
