| US 7,553,620 B2 | ||
| Method for determining polynucleotides in a sample without attaching these to a support, and using detection probes | ||
| Rudolf Rigler, St-Sulpice (Switzerland); and Zeno Foldes-Papp, Graz (Austria) | ||
| Assigned to Gnothis Holdings S.A., Ecublens (Switzerland) | ||
| Appl. No. 10/276,039 PCT Filed May 11, 2001, PCT No. PCT/EP01/05410 § 371(c)(1), (2), (4) Date Mar. 26, 2003, PCT Pub. No. WO01/85991, PCT Pub. Date Nov. 15, 2001. |
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| Claims priority of application No. 100 23 421 (DE), filed on May 12, 2000; and application No. 100 65 632 (DE), filed on Dec. 29, 2000. | ||
| Prior Publication US 2003/0165929 A1, Sep. 04, 2003 | ||
| Int. Cl. C12Q 1/68 (2006.01) | ||
| U.S. Cl. 435—6 | 15 Claims |
| 1. A method for determining a target polynucleotide, comprising:
providing a support containing a plurality of separate detection regions, each of which contains a target polynucleotide to
be determined which is neither directly nor indirectly bound to said support;
contacting the target polynucleotide located in each of said detection regions with a labeled detection probe, with individual
detection regions each containing different combinations of hybridizing target polynucleotide and labeled detection probe
to form hybrid molecules between said target polynucleotide and said labeled detection probe; and
determining hybridization of said target polynucleotide with said labeled detection probe by detecting reduced mobility of
hybrid molecules containing target polynucleotide and labeled detection probe, compared with non-hybrid molecules, wherein
said target polynucleotide is coupled to a non-immobilized microparticle having a size of ≤1 μm, wherein said non-immobilized
microparticle remains in solution during determination of hybridization.
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