Identification and quantification of a plurality of biological (micro)organisms or their components
First Claim
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1. A diagnostic and/or quantification kit of a biological organism or part of an organism comprising:
- (a) specific primers for the amplification of a nucleotide sequence;
(b) a fluorescent label for labelling the amplified sequence;
(c) an insoluble solid support surface upon which single-stranded unlabeled capture molecules are covalently bound, said capture molecules being disposed upon the surface of the solid support according to an array, wherein said capture molecules comprise a spacer portion comprising a nucleotide sequence of at least 40 bases in length and a capture portion of between 10 and 60 bases which is able to specifically bind to a nucleotide sequence of said organism or part of it; and
(d) a closed reaction chamber for performing a genetic amplification together with the identification and/or quantification of amplified nucleotide sequences from said organism or part of it.
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Abstract
Disclosed is a system and method conducting real-time PCR. Capture molecules of a specific design are immobilized on a solid support, and contacted with amplicons produced in one or more PCR cycles. Detection of amplicons may take place during or between the PCR cycles while the solid support is in fluidic contact with the PCR solution. In an alternate embodiment detection of the amplicons takes place when the solid support is not in fluidic contact with the PCR solution. The method is suitable for the simultaneous detection and quantification of closely homologous target molecules.
48 Citations
14 Claims
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1. A diagnostic and/or quantification kit of a biological organism or part of an organism comprising:
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(a) specific primers for the amplification of a nucleotide sequence; (b) a fluorescent label for labelling the amplified sequence; (c) an insoluble solid support surface upon which single-stranded unlabeled capture molecules are covalently bound, said capture molecules being disposed upon the surface of the solid support according to an array, wherein said capture molecules comprise a spacer portion comprising a nucleotide sequence of at least 40 bases in length and a capture portion of between 10 and 60 bases which is able to specifically bind to a nucleotide sequence of said organism or part of it; and (d) a closed reaction chamber for performing a genetic amplification together with the identification and/or quantification of amplified nucleotide sequences from said organism or part of it. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13)
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14. A diagnostic and/or quantification kit of a biological organism or part of an organism comprising:
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(a) specific primers for the amplification of a nucleotide sequence; (b) a fluorescent label for labelling the amplified sequence; (c) an insoluble solid support surface upon which single-stranded unlabeled capture molecules are covalently bound, said capture molecules being disposed upon the surface of the solid support according to an array, wherein said capture molecules comprise a spacer portion comprising a nucleotide sequence of at least 40 bases in length and a capture portion of between 10 to 60 bases which is able to specifically bind to a nucleotide sequence of said organism or part of it; (d) a closed reaction chamber for performing a genetic amplification together with the identification and/or quantification of amplified nucleotide sequences from said organism or part of it; and (e) a microlaboratory system based upon the use of microfluidic technology which is able to perform the genetic amplification and the identification of amplified nucleotide sequence upon the same solid support.
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Specification