Fast results hybrid capture assay and system

• US 8,735,564 B2
• Filed: 09/14/2012
• Issued: 05/27/2014
• Est. Priority Date: 10/27/2008
• Status: Active Grant

• Current Assignee: QIAGEN Gaithersburg, Inc. (Qiagen NV)
• Original Assignee: QIAGEN Gaithersburg, Inc. (Qiagen NV)
• Inventors: Eder, Paul, Nazarenko, Irina, Ramachandran, Suganthi, Bell, Laura, Payne, Eric, Virmani, Arvind
• Primary Examiner(s): Martinell, James
• Application Number:
  US13/616,899
• Publication Number:
  US 20130011830A1
• Time in Patent Office: 620 Days
• Field of Search: None
• US Class Current: 536/24.3
• CPC Class Codes: C12Q 1/6804 Nucleic acid analysis using immunogens immunoassay G01N33/53
• CPC Class Codes: C12Q 1/708 for papilloma
• CPC Class Codes: C12Q 2527/125 Specific component of sample, medium or buffer
• CPC Class Codes: C12Q 2527/153 Viscosity

Abstract

The present invention comprises a method that provides fast and reliable results for detecting the presence of a target nucleic acid molecule in a sample.

Claims

1. A composition comprising:

(a) a biological sample suspended in a collection medium, wherein said collection medium comprises NP-40, sodium deoxycholate, and EDTA;

(b) a denaturation reagent;

(c) at least one polynucleotide probe capable of binding to a target nucleic acid molecule;

(d) a support coated with a first antibody; and

(e) a second antibody labeled with a detectable marker.

2. The composition of claim 1, wherein said biological sample is a cervical cell sample.

3. The composition of claim 1, wherein said collection medium comprises about 0.8% to about 1.5% NP-40, about 0.20% to about 0.40% sodium deoxycholate, and about 20 mM to about 40 mM EDTA.

4. The composition of claim 1, wherein a RLU/CO value for said biological sample does not change with time during at least a 21 day period when stored in said collection medium at 33°

C.

5. The composition of claim 1, wherein said at least one polynucleotide probe is selected from the group consisting of probes for HPV high risk types 16, 18, 26, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, 68, and 82.

6. The composition of claim 1, wherein said collection medium further comprises sodium azide.

7. A composition comprising:

(a) a biological sample suspended in a collection medium comprising NP-40, sodium deoxycholate, and EDTA; and

(b) a polynucleotide probe.

8. The composition of claim 7, wherein said collection medium further comprises sodium azide.

9. The composition of claim 7, wherein said biological sample is a cervical cell sample.

10. The composition of claim 7, wherein a RLU/CO value for said biological sample does not change with time during at least a 21 day period when stored in said collection medium at 33°

C.

11. The composition of claim 7, wherein nucleic acid molecules in said biological sample are denatured.

12. The composition of claim 7, further comprising:

(c) a support coated with a first antibody.

13. The composition of claim 12, further comprising:

(d) a second antibody.

14. The composition of claim 13, wherein said second antibody is labeled with a detectable marker.

15. A kit for the detection of a target nucleic acid molecule in a sample, comprising,

a) a collection medium comprising NP-40, sodium deoxycholate, and EDTA;

b) a denaturation reagent;

c) a support coated with a first anti-hybrid antibody;

d) a detection reagent comprising a second anti-poly hybrid antibody, wherein the second antibody is detectably labeled;

e) a detergent-based wash buffer; and

f) a second detection reagent comprising a substrate for the label on the second antibody.

16. The kit of claim 15, wherein said collection medium further comprises sodium azide.

17. The kit of claim 15, wherein said detergent-based wash buffer comprises about 0.8% to about 1.5% NP-40, about 0.20% to about 0.40% sodium deoxycholate, about 30 mM to about 60 mM Tris-HCl, about 20 mM to about 40 mM EDTA, about 100 mM to about 200 mM NaCl, and sodium azide.

18. The kit of claim 15, wherein said detergent-based wash buffer comprises about 40 mM Tris pH 8.2, 100 mM NaCl, 0.1% - 0.5% Triton x-100, and 0.05% sodium azide.

19. The kit of claim 15, further comprising a probe diluent, wherein the diluent comprises BES, polyacrylic acid, NaOH and sodium azide.

20. The kit of claim 15, wherein the denaturation reagent is 1.75 N NaOH.

21. The kit of claim 15, further comprising at least one polynucleotide probe capable of binding to human papillomavirus (HPV) and genetic variants thereof.

22. The kit of claim 21, wherein said at least one polynucleotide probe is selected from the group consisting of probes for HPV high risk types 16, 18, 26, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, 68,and 82.

23. The kit of claim 15, further comprising an apparatus configured to detect the presence of a target nucleic acid in a sample.

24. The kit of claim 23, wherein said system is a luminometer.