Method for detecting the presence of a nucleic acid in a sample
View Patent ↗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.
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 in the sample, 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 complex contained in the sample to a magnetic field;
c) while the complex contained in the sample is subjected to the magnetic field, aspirating at least a portion of the fluid component of the sample from the complex;
d) after step c), washing the solid support one or more times with a wash buffer, thereby providing a purified form of the nucleic acid;
e) forming a reaction mixture with a pipette of the analyzer, wherein the reaction mixture comprises the purified form of the nucleic acid and all reagents required to perform a nucleic acid amplification;
f) synthesizing amplification products in the reaction mixture, each of the amplification products comprising a nucleotide sequence contained in the nucleic acid or its complement;
g) exposing an amplification product that is one of the amplification products synthesized in step f) to a probe having a label, such that a hybrid comprising the probe and the amplification product is formed in solution in a mixture containing the probe and the amplification products; and
h) in the mixture of step g), detecting the label after 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,
wherein steps b)-d) are performed at a first station of the analyzer and step f) is performed at a second station of the analyzer, and
wherein step f) is performed within an enclosure having a receptacle access opening, the receptacle access opening being closed by a door of the enclosure during step f).
2. The method of claim 1 , further comprising, prior to step a), a manual step of providing the sample and the reagents to the analyzer, wherein the sample and the reagents are separately provided to the analyzer.
3. The method of claim 1 , wherein the complex formed in step a) further comprises a capture probe hybridized to the nucleic acid.
4. The method of claim 1 , wherein the fluid component is aspirated through a tiplet in frictional engagement with an aspirator tube of a fluid aspirator of the analyzer.
5. The method of claim 1 , further comprising, prior to step c), an automated step of sensing the level of the fluid component.
6. The method of claim 1 , wherein the portion of the fluid component aspirated in step c) is drawn through an aspirator tube to a fluid waste container.
7. The method of claim 6 , wherein the fluid waste container is situated in a lower chassis of the analyzer beneath a processing deck contained within the housing.
8. The method of claim 1 , wherein the steps of the method are performed in a single receptacle.
9. The method of claim 8 , wherein the receptacle is a cylindrical tube.
10. The method of claim 8 , wherein the receptacle is one of a plurality of integrally formed receptacles.
11. The method of claim 1 , wherein the amplification products formed in step f) comprise amplification products having the nucleotide sequence contained in the nucleic acid and amplification products having the nucleotide sequence contained in the complement of the nucleic acid.
12. The method of claim 1 , wherein the probe hybridizes with specificity to the amplification product in step g), and wherein the nucleic acid is from a disease-associated pathogen.
13. The method of claim 1 , wherein the label is a fluorescent or chemiluminescent label.
14. The method of claim 1 , wherein the first and second stations are situated on a processing deck contained within the housing.
15. The method of claim 1 , wherein steps f) and g) are performed at separate stations situated within the housing.
16. The method of claim 15 , wherein step f) is performed in a first incubator and step g) is performed in a second incubator, and wherein the first and second incubators are in a laterally spaced-apart relationship.
17. The method of claim 15 , wherein the stations are situated on a processing deck contained within the housing.
18. The method of claim 1 , wherein the analyzer is controlled by a computer controller that is integrally housed within the analyzer.
19. The method of claim 1 , wherein the steps of the method are performed at multiple stations situated on a processing deck contained within the housing.
20. The method of claim 1 , wherein the housing remains closed when the steps of the method are performed.
21. The method of claim 1 , wherein step f) is performed at a substantially uniform temperature.