Claims
- 1. An automated method for determining whether a target nucleic acid is present in a sample contained by an open-ended receptacle vessel, said method comprising the steps of:
a) separating said target nucleic acid, if present in said sample, from non-target nucleic acid which may be present in said sample; b) withdrawing at least a portion of said sample to remove non-target nucleic which may be present in said sample from said receptacle vessel; c) exposing the contents of said receptacle vessel to reagents and conditions sufficient to amplify a target nucleic acid sequence contained within said target nucleic acid; and d) determining whether said target nucleic acid sequence has been amplified as an indication of the presence or absence of said target nucleic acid in said sample, wherein steps a)-d) are automated and performed on a single platform.
- 2. The method of claim 1, wherein said separating step includes providing to said sample a support material for immobilizing said target nucleic acid.
- 3. The method of claim 2, wherein said support material comprises magnetically-responsive particles.
- 4. The method of claim 3, wherein said separating step includes subjecting the contents of said receptacle vessel to a magnetic field, thereby isolating said magnetically-responsive particles within said receptacle vessel.
- 5. The method of claim 4, wherein said separating step further includes providing to said sample a capture probe having a first nucleotide base sequence region which hybridizes to said target nucleic acid under a first set of predetermined hybridization conditions.
- 6. The method of claim 5, wherein said capture probe has a second nucleotide base sequence region which hybridizes to a polynucleotide immobilized by said magnetically-responsive particles under a second set of predetermined hybridization conditions, said first and second sets of predetermined hybridization conditions being different.
- 7. The method of claim 4, wherein said withdrawing step includes aspirating at least a portion of said sample from said receptacle vessel after isolating said magnetically-responsive particles within said receptacle vessel.
- 8. The method of claim 7 further comprising the step of washing the contents of said receptacle vessel with a wash buffer between said withdrawing and exposing steps.
- 9. The method of claim 1, wherein said determining step includes providing a detection probe containing a nucleotide base sequence to the contents of said receptacle vessel under conditions such that said detection probe specifically hybridizes to nucleic acid in said receptacle vessel which contains said target sequence or the complement thereof.
- 10. The method of claim 9, wherein said detection probe includes a label.
- 11. The method of claim 10, wherein said label is selected from the group consisting of a radiolabel, a fluorescent dye, biotin, an enzyme and a chemiluminescent compound.
- 12. The method of claim 9, wherein said determining step provides an indication of the presence or absence of at least one member of a target group of organisms or viruses.
- 13. The method of claim 1, wherein said receptacle vessel is a component of an integral reaction receptacle comprising multiple receptacle vessels.
- 14. The method of claim 1, wherein said platform is contained within a housing of an analyzer.
- 15. The method of claim 14, wherein said analyzer is moveable.
- 16. An automated method for determining whether a target nucleic acid is present in a sample contained by an open-ended receptacle vessel, said method comprising the steps of:
a) providing a support material to said receptacle vessel, said support material comprising magnetically-responsive particles for immobilizing said target nucleic acid; b) subjecting the contents of said receptacle vessel to a magnetic field, thereby isolating said magnetically-responsive particles within said receptacle vessel; c) aspirating at least a portion of said sample from said receptacle vessel, thereby removing non-target nucleic acid present in said sample from said receptacle vessel; d) washing the contents of said receptacle vessel with a wash buffer; e) exposing the contents of said receptacle vessel to reagents and conditions sufficient to amplify a target nucleic acid sequence contained within said target nucleic acid; and f) determining whether said target nucleic acid sequence has been amplified as an indication of the presence or absence of said target nucleic acid in said sample, wherein steps a)-f) are automated and performed on a single platform.
- 17. The method of claim 16, wherein said separating step further includes providing to said sample a capture probe having a first nucleotide base sequence region which hybridizes to said target nucleic acid under a first set of predetermined hybridization conditions.
- 18. The method of claim 17, wherein said capture probe has a second nucleotide base sequence region which hybridizes to a polynucleotide immobilized by said magnetically-responsive particles under a second set of predetermined hybridization conditions, said first and second sets of predetermined hybridization conditions being different.
- 19. The method of claim 16, wherein said determining step includes providing a detection probe containing a nucleotide base sequence to the contents of said receptacle vessel under conditions such that said detection probe specifically hybridizes to nucleic acid in said receptacle vessel which contains said target sequence or the complement thereof.
- 20. The method of claim 19, wherein said detection probe includes a label.
- 21. The method of claim 20, wherein said label is selected from the group consisting of a radiolabel, a fluorescent dye, biotin, an enzyme and a chemiluminescent compound.
- 22. The method of claim 19, wherein said determining step provides an indication of the presence or absence of at least one member of a target group of organisms or viruses.
- 23. The method of claim 16, wherein said receptacle vessel is a component of an integral reaction receptacle comprising multiple receptacle vessels.
- 24. The method of claim 16, wherein said platform is contained within a housing of an analyzer.
- 25. The method of claim 24, wherein said analyzer is moveable.
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation of U.S. patent application Ser. No. 09/985,064 filed Nov. 1, 2001, pending, which is a continuation of U.S. patent application Ser. No. 09/303,030 filed Apr. 30, 1999, now U.S. Pat. No. 6,335,166, which claims the benefit of U.S. Provisional Application No. 60/083,927 filed May 1, 1998. The disclosure of U.S. patent application Ser. No. 09/985,064 is hereby incorporated herein by reference.
Provisional Applications (1)
|
Number |
Date |
Country |
|
60083927 |
May 1998 |
US |
Continuations (2)
|
Number |
Date |
Country |
Parent |
09985064 |
Nov 2001 |
US |
Child |
10263939 |
Oct 2002 |
US |
Parent |
09303030 |
Apr 1999 |
US |
Child |
09985064 |
Nov 2001 |
US |