Method for detecting the presence of a nucleic acid in a sample
First Claim
1. A method for detecting the presence of a nucleic acid in a sample, the method comprising performing within a housing of a self-contained, stand-alone analyzer the automated steps of:
- a) contacting the sample with a solid support such that a complex comprising the nucleic acid and the solid support is formed, wherein the solid support comprises a magnetically-responsive particle, and wherein the complex is suspended in a fluid component of the sample;
b) after step a), subjecting the sample containing the complex to a magnetic field;
c) while the sample containing the complex is subjected to the magnetic field, aspirating at least a portion of the fluid component of the sample from the complex, thereby purifying the nucleic acid;
d) forming a reaction mixture with a pipette of the analyzer, wherein the reaction mixture comprises the nucleic acid purified in step c) and all reagents required to perform a nucleic acid amplification;
e) synthesizing amplification products in the reaction mixture, each of the amplification products comprising a nucleotide sequence contained in the nucleic acid or its complement;
f) exposing an amplification product that is one of the amplification products synthesized in step e) to a probe, such that a hybrid comprising the probe and the amplification product is formed in a mixture containing the probe and the amplification products; and
g) detecting the formation of the hybrid, wherein the formation of the hybrid in the mixture is an indication of the presence of the nucleic acid in the sample.
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Abstract
An automated method for detecting the presence of a nucleic acid in a sample, where the method is performed within a housing of a self-contained, stand-alone analyzer. The method includes purifying the nucleic acid after it has been immobilized on a magnetically-responsive solid support. A pipette of the analyzer is used to form a reaction mixture comprising the purified nucleic acid and all reagents required to perform a nucleic acid amplification. Amplification products are synthesized that include a nucleotide sequence contained in the nucleic acid or the complement of the nucleic acid. The amplification products are exposed to a probe in a mixture, where the probe forms a hybrid with one of the amplification products. The formation of the hybrid in the mixture provides an indication of the presence of the nucleic acid in the sample.
379 Citations
27 Claims
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1. A method for detecting the presence of a nucleic acid in a sample, the method comprising performing within a housing of a self-contained, stand-alone analyzer the automated steps of:
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a) contacting the sample with a solid support such that a complex comprising the nucleic acid and the solid support is formed, wherein the solid support comprises a magnetically-responsive particle, and wherein the complex is suspended in a fluid component of the sample; b) after step a), subjecting the sample containing the complex to a magnetic field; c) while the sample containing the complex is subjected to the magnetic field, aspirating at least a portion of the fluid component of the sample from the complex, thereby purifying the nucleic acid; d) forming a reaction mixture with a pipette of the analyzer, wherein the reaction mixture comprises the nucleic acid purified in step c) and all reagents required to perform a nucleic acid amplification; e) synthesizing amplification products in the reaction mixture, each of the amplification products comprising a nucleotide sequence contained in the nucleic acid or its complement; f) exposing an amplification product that is one of the amplification products synthesized in step e) to a probe, such that a hybrid comprising the probe and the amplification product is formed in a mixture containing the probe and the amplification products; and g) detecting the formation of the hybrid, wherein the formation of the hybrid in the mixture is an indication of the presence of the nucleic acid in the sample. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27)
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Specification