Patent ID: 6261771
Filing Date: 2001-07-17
Classification: C12Q

Abstract:
A method for detecting the presence of extremely low levels of multiple target nucleic acid sequences and/or multiple target antigens in a sample, comprising:(a) presenting a sample suspected of containing targets selected from the group consisting of nucleic acids, antigens, and mixtures of at least one nucleic acid and at least one antigen, to a plurality of collector molecules comprised of a first complementary molecule and a second complementary molecule, the first and second complementary molecules being hybridized and bound to one another, the first complementary molecule being attached to a support medium and the second complimentary molecule comprises a complimentary binding portion and a unique reporter oligonucleotide molecule flanked by primer binding sites such that the targets in the sample displace the second complementary molecule which is bound to the first complementary molecule and hybridize with the first complementary molecule thereby releasing the second complementary molecule into the sample;(b) washing the sample containing the released second complementary molecule;(c) mixing the sample containing the released second complementary molecule with labeled primers and polymerase chain reaction reagents;(d) amplifying the unique reporter oligonucleotide molecule and flanking primer binding sites of the second complementary molecule by passing the sample through a conduit in intimate contact with a surface having a temperature gradient suitable for amplification or reaction conditions;(e) presenting the sample containing amplified unique reporter nucleic acid molecules to a plurality of different binding sites arranged in a known order, with respect to each other, wherein a third molecule complementary to the amplified unique reporter molecule is bound to at least one of the collector binding sites; and(f) detecting at least one collector binding site for a light emission indicative of the presence of an amplified and labeled unique reporter molecule bound to the third complementary molecule.