Integrated nucleic acid analysis
First Claim
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1. A method for analyzing a target nucleic acid in a biological sample, comprising the following steps:
- a) providing a reaction vial consisting of a single compartment and a closable lid, wherein said reaction vial contains;
i) an absorbent matrix;
ii) a biological sample on said absorbent matrix, wherein said biological sample comprises said target nucleic acid, and wherein said nucleic acid has not been extracted, washed, and/or eluted from said biological sample;
iii) a reagent mixture for amplifying said nucleic acid;
iv) a reagent mixture for detecting the amplified nucleic acid, andb) closing the lid of said reaction vial;
c) releasing said nucleic acid from said absorbent matrix;
d) amplifying said nucleic acid; and
e) detecting the amplified nucleic acid through the use of a labelled probe as a result of fluorescence resonance energy transfer;
wherein;
steps c) through e) are performed in said reaction vial without removing said absorbent matrix from said reaction vial and without opening said reaction vial.
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Abstract
The present invention relates to an integrated method of nucleic acid analysis, and more particularly to a simplified sample pre-treatment, which renders the method more easily automated, where the sample is provided on or applied onto a solid matrix and the subsequent amplification and detection steps are performed in one single, sealed reaction vial without removing the matrix.
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Citations
27 Claims
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1. A method for analyzing a target nucleic acid in a biological sample, comprising the following steps:
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a) providing a reaction vial consisting of a single compartment and a closable lid, wherein said reaction vial contains; i) an absorbent matrix; ii) a biological sample on said absorbent matrix, wherein said biological sample comprises said target nucleic acid, and wherein said nucleic acid has not been extracted, washed, and/or eluted from said biological sample; iii) a reagent mixture for amplifying said nucleic acid; iv) a reagent mixture for detecting the amplified nucleic acid, and b) closing the lid of said reaction vial; c) releasing said nucleic acid from said absorbent matrix; d) amplifying said nucleic acid; and e) detecting the amplified nucleic acid through the use of a labelled probe as a result of fluorescence resonance energy transfer;
wherein;steps c) through e) are performed in said reaction vial without removing said absorbent matrix from said reaction vial and without opening said reaction vial. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 27)
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14. A method for analyzing a target nucleic acid in one or more biological samples, comprising the following steps:
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a) providing a multi-well plate, wherein each well of said multi-well plate consists of a single compartment, and wherein one or more wells of said multi-well plate contain; i) an absorbent matrix; ii) a biological sample on said absorbent matrix, wherein said biological sample comprises said target nucleic acid, and wherein said nucleic acid has not been extracted, washed, and/or eluted from said biological sample; iii) a reagent mixture for amplifying said nucleic acid; and iv) a reagent mixture for detecting the amplified nucleic acid; b) sealing said one or more wells of said multi-well plate; c) releasing said nucleic acid from said absorbent matrix; d) amplifying said nucleic acid; and e) detecting the amplified nucleic acid through the use of a labelled probe as a result of fluorescence resonance energy transfer;
wherein;steps c) through e) are performed in said one or more wells of said multi-well plate without removing said absorbent matrix from said one or more wells of said multi-well plate and without opening said one or more wells of said multi-well plate. - View Dependent Claims (15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26)
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Specification