Automated process for detecting the presence of a target nucleic acid in a sample
View Patent ↗An automated process for detecting the presence of a target nucleic acid in a sample. The process includes contacting a sample with a magnetically-responsive solid support and immobilizing a target nucleic acid present in the sample on the solid support. At a first station of an analyzer, the solid support is subjected to a magnetic field and the immobilized target nucleic acid is separated from a non-immobilized component of the sample. A solution comprising the separated target nucleic acid is moved to an incubator of a second station of the analyzer, where the separated target nucleic acid is subjected to an amplification procedure and an amplification product is formed. The amplification product is detected as an indication of the presence of the target nucleic acid in the sample.
1. A process for detecting the presence of a target nucleic acid in a sample, the process comprising the automated steps of:
(a) contacting a sample containing a target nucleic acid with a solid support comprising a magnetically-responsive particle;
(b) immobilizing the target nucleic acid on the solid support;
(c) at a first station of an analyzer, subjecting the solid support to a magnetic field and separating the immobilized target nucleic acid from a non-immobilized component of the sample;
(d) moving a solution comprising the separated target nucleic acid to a second station of the analyzer;
(e) in an incubator of the second station, subjecting the separated target nucleic acid to an amplification procedure and forming an amplification product; and
(f) detecting the amplification product formed in step (e) as an indication of the presence of the target nucleic acid in the sample.
2. The process of claim 1 , wherein step (c) further comprises aspirating the non-immobilized component of the sample while the solid support is subjected to the magnetic field.
3. The process of claim 2 , wherein step (c) further comprises washing the solid support one or more times with a wash buffer after aspirating the non-immobilized component of the sample.
4. The process of claim 1 further comprising hybridizing a capture probe to the target nucleic acid prior to step (b).
5. The process of claim 1 , wherein the non-immobilized component of the sample comprises non-target nucleic acid.
6. The process of claim 1 , wherein the solution is formed in a reaction receptacle.
7. The process of claim 6 , wherein the reaction receptacle is one of an integrally formed plurality of reaction receptacles.
8. The process of claim 6 , wherein the reaction receptacle has an open top end.
9. The process of claim 8 , wherein the reaction receptacle comprises a cylindrical tube.
10. The process of claim 1 , wherein the amplification product is a nucleic acid sequence that is the same as or complementary to a sequence of the target nucleic acid.
11. The process of claim 10 further comprising hybridizing a probe to the amplification product, wherein the amplification product is detected in step (f) by detecting the probe hybridized to the amplification product.
12. The process of claim 11 , wherein the probe includes a detectable label.
13. The process of claim 10 , wherein the amplification reaction is selected from the group consisting of a polymerase chain reaction, a strand displacement amplification, and a transcription-associated amplification.
14. The process of claim 1 , wherein the target nucleic acid is derived from an organism or virus.
15. The process of claim 1 , wherein each of the first and second stations is located on a processing deck of the analyzer.
16. The process of claim 1 , wherein the first and second stations are contained within a housing of the analyzer.
17. The process of claim 1 , wherein each of the steps of the method is performed within the analyzer.
18. A process for detecting the presence of a target nucleic acid in a sample, the process comprising the automated steps of:
(a) contacting a sample containing a target nucleic acid with a solid support comprising a magnetically-responsive particle;
(b) immobilizing the target nucleic acid on the solid support;
(c) at a first station of an analyzer, subjecting the solid support to a magnetic field and separating the immobilized target nucleic acid from a non-immobilized component of the sample;
(d) forming a solution comprising the separated target nucleic acid and an amplification reagent in a reaction receptacle;
(e) moving the reaction receptacle to a second station of the analyzer;
(f) in an incubator of the second station, subjecting the separated target nucleic acid to an amplification procedure and forming an amplification product; and
(g) detecting the amplification product formed in step (f) as an indication of the presence of the target nucleic acid in the sample.
19. The process of claim 18 , wherein the first and second stations are contained within a housing of the analyzer.
20. The process of claim 18 , wherein step (c) comprises aspirating the non-immobilized component of the sample while the magnetically-responsive particle is subjected to the magnetic field.
21. The process of claim 20 , wherein step (c) further comprises washing the solid support one or more times with a wash buffer after aspirating the non-immobilized component of the sample.
22. The process of claim 18 further comprising hybridizing a capture probe to the target nucleic acid prior to step (b).
23. The process of claim 18 , wherein the non-immobilized component of the sample comprises non-target nucleic acid.
24. The process of claim 18 , wherein the reaction receptacle is one of an integrally formed plurality of reaction receptacles.
25. The process of claim 18 , wherein the reaction receptacle has an open top end.
26. The process of claim 25 , wherein the reaction receptacle comprises a cylindrical tube.
27. The process of claim 18 , wherein the reaction receptacle is moved between the first and second stations with a transport mechanism of the analyzer, wherein the transport mechanism comprises a receptacle carrier assembly constructed to be rotatable about an axis of rotation.
28. The process of claim 18 further comprising the automated step of adjusting the temperature of the contents of the reaction receptacle to the approximate temperature of the incubator prior to receiving the reaction receptacle in the incubator.
29. The process of claim 18 further comprising the automated step of dispensing a layer of oil into the reaction receptacle prior to step (f).
30. The process of claim 18 , wherein the amplification product is a nucleic acid sequence that is the same as or complementary to a sequence of the target nucleic acid.
31. The process of claim 30 further comprising hybridizing a probe to the amplification product, wherein the amplification product is detected in step (g) by detecting the probe hybridized to the amplification product.
32. The process of claim 31 , wherein the probe includes a detectable label.
33. The process of claim 30 , wherein the amplification reaction is selected from the group consisting of a polymerase chain reaction, a strand displacement amplification, and a transcription-associated amplification.
34. The process of claim 18 , wherein the target nucleic acid is derived from an organism or virus.
35. The process of claim 18 , wherein each of the first and second stations is located on a processing deck of the analyzer.
36. The process of claim 18 , wherein each of the steps of the method is performed within the analyzer.