Capture and detection format versatility for dipstick assays

• US 7,192,701 B2
• Filed: 07/06/2001
• Issued: 03/20/2007
• Est. Priority Date: 07/07/2000
• Status: Expired due to Term

• Current Assignee: Diagnostics for the Real World, Ltd.
• Original Assignee: Diagnostics for the Real World, Ltd.
• Inventors: Dineva, Magda Anastassova, Lee, Helen, Hu, Hsiang Yun
• Primary Examiner(s): WHISENANT, ETHAN C
• Application Number:
  US10/332,131
• Publication Number:
  US 20040053255A1
• Time in Patent Office: 2,083 Days
• Field of Search: 435/6, 536/23.1, 536/24.3
• US Class Current: 435/6.11
• CPC Class Codes: C12Q 1/6834 Enzymatic or biochemical coupling of nucleic acids to a solid phase
• CPC Class Codes: C12Q 1/689 for bacteria
• CPC Class Codes: C12Q 2565/625 being a nucleic acid test strip device, e.g. dipsticks, strips, tapes, CD plates

Abstract

Dipsticks for testing for the presence of a target nucleic acid in a sample solution are described the dipsticks comprise a universal capture probe immobilised at a capture zone of the dipstick. The universal capture probe is capable of hybridising to a hook capture probe which is hybridised to the target nucleic acid in the sample solution. A contact end of the dipstick is contacted with the sample solution to cause hook capture probe hybridised to the target nucleic acid to move by capillary action to the capture zone where the target nucleic acid can be detected. Use of the universal and hook capture probes allows dipsticks to be prepared which can be used to capture any target nucleic acid, thereby simplifying preparation of the dipsticks. Specificity of target nucleic acid capture is then achieved by use of an appropriate hook capture probe. Methods and kits are also described.

Claims

1. A dipstick for testing for the presence of a target nucleic acid in a sample solution, wherein the dipstick comprises a chromatographic strip having:

a contact end for contacting the sample solution;

a universal capture probe that is immobilized at a capture zone of the chromatographic strip remote from the contact end; and

a hook capture probe hybridized to the universal capture probe and which is capable of hybridizing to a first region of the target nucleic acid.

3. The dipstick according to claim 1 further comprises a detection moiety that is releasably immobilized at a conjugate zone of the chromatographic strip between the contact end and the capture zone, wherein the detection moiety is capable of binding by non base pairing interaction to a detection probe that is hybridized to a second region of the target nucleic acid.

15. The dipstick of claim 1, wherein the target nucleic acid is nucleic acid of a disease causing microorganism, wherein the microorganism is a sexually transmitted disease causing microorganism.

16. The dipstick according to claim 15, wherein the target nucleic acid is Chlamydia trachomatis nucleic acid.

17. A method of testing for the presence of a target nucleic acid in a sample solution comprising contacting the dipstick of claim 1 with the sample solution suspected of comprising the target nucleic acid.

2. A dipstick for testing for the presence of a target nucleic acid in a sample solution, wherein the dipstick comprises a chromatographic strip having;

a contact end for contacting the sample solution;

a universal capture probe that is immobilized at a capture zone of the chromatographic strip remote from the contact end; and

a hook capture probe that is releasably immobilized to the chromatographic strip between the contact end and the capture zone, wherein the hook capture probe is capable of hybridizing to a first region of the target nucleic acid, and the universal capture probe is capable of hybridizing to the hook capture probe.

4. A dipstick for testing for the presence of a target nucleic acid in a sample solution, wherein the dipstick comprises a chromatographic strip having:

a contact end for contacting the sample solution;

a universal capture probe that is immobilized at a capture zone of the chromatographic strip remote from the contact end, wherein the universal capture probe is capable of hybridizing to a hook capture probe bound to a first region of the target nucleic acid; and

a detection moiety that is releasably immobilized at a conjugate zone of the chromatographic strip between the contact end and the capture zone, wherein the detection moiety is capable of binding by non base pairing interaction to a detection probe hybridized to a second region of the target nucleic acid.

5. The dipstick according to claim 4, wherein the detection moiety comprises a label.

6. The dipstick according to claim 5, wherein the label is non radioactive.

7. The dipstick according to claim 6, wherein the label is a color label.

8. The dipstick according to claim 7, wherein the color label comprises a textile dye, a metal sol, or a colored particle.

9. The dipstick according to claim 4, wherein the detection moiety comprises an antibody or an antibody fragment.

10. The dipstick according to claim 9, wherein the detection moiety comprises an anti-biotin antibody, an anti-fluorescein antibody, or an anti-DNP antibody.

11. The dipstick according to claim 4, wherein the detection moiety comprises avidin, streptavidin, or a derivative thereof which retains biotin binding activity.

12. A dipstick for testing for the presence of a target nucleic acid in a sample solution, wherein the dipstick comprises a chromatographic strip having:

a contact end for contacting the sample solution;

a capture probe that is immobilized at a capture zone of the chromatographic strip remote from the contact end, wherein the capture probe is capable of binding directly or indirectly to the target nucleic acid; and

a universal detection probe that is releasably immobilized to the chromatographic strip between the contact end and the capture zone, wherein the universal detection probe is capable of hybridizing to a hook detection probe.

13. The dipstick of claim 12 further comprises a detection moiety that is releasably immobilized at a conjugate zone of the chromatographic strip between the contact end and the capture zone, wherein the detection moiety is capable of binding by non base pairing interaction to the universal detection probe.

35. The dipstick of claim 12 further comprises a hook detection probe that is releasably immobilized to the chromatographic strip between the contact end and the capture zone, wherein the hook detection probe is capable of hybridizing to the target nucleic acid.

14. A dipstick for testing for the presence of a target nucleic acid in a sample solution, wherein the dipstick comprises:

a dipstick having a chromatographic strip with a contact end for contacting the sample solution and a capture moiety that is immobilized at a capture zone of the chromatographic strip remote from the contact end, wherein the capture moiety is capable of binding by non base pairing interaction to a universal capture probe bound to the target nucleic acid by a hook capture probe; and

a hook capture probe or a universal capture probe that is releasably immobilized to the chromatographic strip between the contact end and the capture zone, wherein the hook capture probe is capable of hybridizing to the target nucleic acid, and the universal capture probe is capable of hybridizing to the hook capture probe.

18. A kit for testing for the presence of a target nucleic acid in a sample solution, wherein the kit comprises:

i) a dipstick comprising a chromatographic strip with a contact end for contacting the sample solution, and a universal capture probe that is immobilized at a capture zone of the chromatographic strip remote from the contact end; and

ii) a hook capture probe capable of hybridizing to a first region of the target nucleic acid and to the universal capture probe.

34. A method of testing for the presence of a target nucleic acid in a sample solution comprising using the kit of claim 18, wherein a dipstick of the kit is contacted with the sample solution suspected of comprising the target nucleic acid.

37. The kit of claim 18 further comprises a detection probe capable of attaching to the target nucleic acid thereby allowing direct or indirect detection of the target nucleic acid utilizing the detection probe.

19. A kit for testing for the presence of a target nucleic acid in a sample solution, wherein the kit comprises:

a dipstick comprising a chromatographic strip with a contact end for contacting the sample solution, a universal capture probe that is immobilized at a capture zone of the chromatographic strip remote from the contact end, and a hook capture probe hybridized to the universal capture probe, wherein the hook probe is capable of hybridizing to a first region of the target nucleic acid.

38. The kit of claim 19 further comprises a detection probe capable of attaching to the target nucleic acid thereby allowing direct or indirect detection of the target nucleic acid utilizing the detection probe.

20. A kit for testing for the presence of a target nucleic acid in a sample solution, wherein the kit comprises:

i) a dipstick comprising a chromatographic strip with a contact end for contacting the sample solution, and a capture probe that is immobilized at a capture zone of the chromatographic strip remote from the contact end, wherein the capture probe is capable of hybridizing to a first region of the target nucleic acid, or is capable of binding indirectly to the target nucleic acid;

ii) a hook detection probe capable of hybridizing to a second region of the target nucleic add; and

iii) a universal detection probe capable of hybridizing to the hook detection probe when the hook detection probe is hybridized to the second region of the target nucleic acid, thereby allowing direct or indirect detection of the target nucleic acid utilizing the detection probe.

21. The kit according to claim 20, wherein the detection probe or the universal detection probe comprises a label thereby allowing direct detection of target nucleic acid utilizing the detection probe or the universal detection probe and the hook detection probes.

22. The kit according to claim 20, wherein the detection probe or the universal detection probe comprises a detection ligand which can be bound by a detection moiety thereby allowing indirect detection of target nucleic acid utilizing the detection probe or the universal detection probe and the hook detection probes.

23. The kit of claim 22 further comprises a detection moiety capable of binding to the detection ligand.

26. The kit according to claim 23, wherein the detection moiety is releasably immobilized at a conjugate zone located between the contact end and the capture zone of the chromatographic strip.

27. The kit according to claim 23, wherein the detection moiety comprises a label thereby allowing detection of the detection moiety.

28. The kit according to claim 21, wherein the label is non radioactive.

29. The kit according to claim 28, wherein the label is a color label.

30. The kit according to claim 29, wherein the color label comprises a textile dye, a metal sol, or a colored particle.

24. The kit of 23, wherein the detection moiety comprises an antibody or an antibody fragment.

25. The kit of 24, wherein the detection ligand comprises a biotin and the detection moiety comprises an anti-biotin antibody, or wherein the detection ligand comprises a fluorescein and the detection moiety comprises an anti-fluorescein antibody, or wherein the detection ligand comprises a 2,4-dinitrophenol (DNP) and the detection moiety comprises an anti-DNP antibody.

31. A kit for testing for the presence of a target nucleic acid in a sample solution, wherein the kit comprises:

a dipstick having a chromatographic strip with a contact end for contacting the sample solution and a capture moiety that is immobilized at a capture zone of the chromatographic strip remote from the contact end, wherein the capture moiety is capable of binding by non base pairing interaction to a universal capture probe bound to the target nucleic acid by a hook capture probe;

a hook capture probe capable of hybridizing to the target nucleic acid; and

a universal capture probe capable of hybridizing to the hook capture probe.

32. A method of testing for the presence of a target nucleic acid in a sample solution using a dipstick assay, wherein the method comprises contacting a dipstick with the sample solution suspected of comprising the target nucleic acid, wherein the dipstick comprises a universal capture probe and a hook capture probe.

33. A method of testing for the presence of a target nucleic acid in a sample solution using a dipstick assay, wherein the method comprises contacting a dipstick with the sample solution suspected of comprising the target nucleic acid, wherein the dipstick comprises a universal detection probe and a hook detection probe.

36. A dipstick for testing for the presence of a target nucleic acid in a sample solution, wherein the dipstick comprises:

a dipstick having a chromatographic strip with a contact end for contacting the sample solution and a capture moiety that is immobilized at a capture zone of the chromatographic strip remote from the contact end, wherein the capture moiety is capable of binding by non base pairing interaction to a universal capture probe bound to the target nucleic acid by a hook capture probe; and

a hook capture probe and a universal capture probe that is releasably immobilized to the chromatographic strip between the contact end and the capture zone, wherein the hook capture probe is capable of hybridizing to the target nucleic acid, and the universal capture probe is capable of hybridizing to the hook capture probe.