FAST RESULTS HYBRID CAPTURE ASSAY AND SYSTEM

• US 20100159463A1
• Filed: 10/26/2009
• Published: 06/24/2010
• 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, Payne, Eric, Bell, Laura, Nazarenko, Irina, Ramachandran, Suganthi, Virmani, Arvind
• Granted Patent: US 8,288,520 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• 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 at least one detergent;

(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 NP-40, sodium deoxycholate, and EDTA.

4. The composition of claim 1, wherein said collection medium comprises about 0.5% to about 2.0% NP-40, about 0.10% to about 0.40% sodium deoxycholate, and about 10 mM to about 50 mM EDTA.

5. The composition of claim 1, wherein said biological sample is stable when stored in said collection medium for at least 21 days at 33°

C.

6. 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.

7. A composition comprising:

(a) a biological sample suspended in a collection medium comprising about 0.5% to about 2.0% NP-40, about 0.10% to about 0.40% sodium deoxycholate, and about 10 mM to about 50 mM EDTA, and

(b) a polynucleotide probe.

8. The composition of claim 7, wherein said collection medium further comprises about 0.01% to about 0.10% sodium azide.

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

10. The composition of claim 7, wherein said biological sample is stable when stored in said collection medium for at least 21 days 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 a detergent;

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 comprises NP-40, Deoxycholate, and EDTA.

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

18. The kit of claim 15, wherein said detergent-based wash buffer comprises about 0.5% to about 2.0% NP-40, about 0.10% to about 0.40% sodium deoxycholate, about 25 mM to about 75 mM Tris-HCl, about 10 mM to about 50 mM EDTA, about 50 mM to about 200 mM NaCl, and about 0.01% to about 0.10% sodium azide.

19. 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.

20. The kit of claim 15, wherein the diluent comprises BES, polyacrylic acid, NaOH and sodium azide.

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

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

23. The kit of claim 22, 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.

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

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

26. A portable system for detecting the presence of a target nucleic molecule acid in a sample comprising:

(a) a heater configured for heating multiple samples;

(b) a luminometer configured to detect the chemiluminescence of a target nucleic acid molecule on a support coated with a first antibody and conjugated to a second antibody labeled with a detectable marker; and

(c) a monitor configured to report chemiluminescence data.