Capture and detection format versatility for dipstick assays
First Claim
1. A method for testing for the presence of target nucleic acid in a sample solution which comprises:
- a) providing a chromatographic strip having a contact end for contacting the sample solution and a universal capture probe immobilised at a capture zone of the chromatographic strip remote from the contact end;
b) providing a hook capture probe capable of hybridising to a first region of the target nucleic acid and to the universal capture probe;
c) incubating the sample solution with the hook capture probe under conditions for hybridisation of the hook capture probe to the first region of target nucleic acid in the sample solution;
d) contacting the sample solution with the contact end of the chromatographic strip so that a complex formed between the hook capture probe, the detection probe and target nucleic acid can travel by capillary action to the capture zone and bind to the capture zone by interaction between the hook capture probe of the complex and the universal capture probe immobilised at the capture zone; and
e) detecting for target nucleic acid at the capture zone.
1 Assignment
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Accused Products
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.
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Citations
63 Claims
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1. A method for testing for the presence of target nucleic acid in a sample solution which comprises:
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a) providing a chromatographic strip having a contact end for contacting the sample solution and a universal capture probe immobilised at a capture zone of the chromatographic strip remote from the contact end;
b) providing a hook capture probe capable of hybridising to a first region of the target nucleic acid and to the universal capture probe;
c) incubating the sample solution with the hook capture probe under conditions for hybridisation of the hook capture probe to the first region of target nucleic acid in the sample solution;
d) contacting the sample solution with the contact end of the chromatographic strip so that a complex formed between the hook capture probe, the detection probe and target nucleic acid can travel by capillary action to the capture zone and bind to the capture zone by interaction between the hook capture probe of the complex and the universal capture probe immobilised at the capture zone; and
e) detecting for target nucleic acid at the capture zone.
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2. A method for testing for the presence of target nucleic acid in a sample solution which comprises:
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a) providing a chromatographic strip having;
a contact end for contacting the sample solution;
a universal capture probe immobilised at a capture zone of the chromatographic strip remote from the contact end; and
a hook capture probe releasably immobilised to the chromatographic strip between the contact end and the capture zone, the hook capture probe being capable of hybridising to a first region of the target nucleic acid and to the universal capture probe;
b) contacting the sample solution with the contact end of the chromatographic strip to cause sample solution to move by capillary action to the capture zone, thereby releasing the hook capture probe and allowing released hook capture probe to hybridise to target nucleic acid in the sample solution as it moves to the capture zone so that a complex comprising target nucleic acid and hook capture probe can be captured at the capture zone by hybridisation of the universal capture probe to the hook capture probe of the complex; and
c) detecting for target nucleic acid at the capture zone.
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3. A method for testing for the presence of target nucleic acid in a sample solution which comprises:
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a) providing a chromatographic strip having a contact end for contacting the sample solution and a universal capture probe immobilised at a capture zone of the chromatographic strip remote from the contact end;
b) providing a hook capture probe capable of hybridising to a first region of the target nucleic acid and to the universal capture probe;
c) providing a detection probe capable of attaching to the target nucleic acid thereby allowing direct or indirect detection of the target nucleic acid utilising the detection probe;
d) incubating the sample solution with the detection probe under conditions for attachment of the detection probe to target nucleic acid in the sample solution; and
incubating the sample solution with the hook capture probe under conditions for hybridisation of the hook capture probe to the first region of target nucleic acid in the sample solution;
e) contacting the sample solution with the contact end of the chromatographic strip so that a complex formed between the hook capture probe, the detection probe and target nucleic acid can travel by capillary action to the capture zone and bind to the capture zone by interaction between the hook capture probe of the complex and the universal capture probe immobilised at the capture zone; and
f) checking for the presence of detection probe at the capture zone. - View Dependent Claims (6, 7, 8, 9, 10, 11, 13, 14, 15, 16, 17)
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4. A method for testing for the presence of target nucleic acid in a sample solution which comprises:
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a) providing a chromatographic strip having a contact end for contacting the sample solution, a universal capture probe immobilised at a capture zone of the chromatographic strip remote from the contact end, and a hook capture probe hybridised to the universal capture probe, the hook probe being capable of hybridising to a first region of the target nucleic acid;
b) contacting the sample solution with the contact end of the chromatographic strip so that target nucleic acid in the sample solution can travel by capillary action to the capture zone and bind to the capture zone by interaction between the hook capture probe immobilised at the capture zone and the first region of the target nucleic acid; and
c) detecting for target nucleic acid at the capture zone.
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5. A method for testing for the presence of target nucleic acid in a sample solution which comprises:
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a) providing a chromatographic strip having a contact end for contacting the sample solution, a universal capture probe immobilised at a capture zone of the chromatographic strip remote from the contact end, and a hook capture probe hybridised to the universal capture probe, the hook probe being capable of hybridising to a first region of the target nucleic acid;
b) providing a detection probe capable of attaching to the target nucleic acid thereby allowing direct or indirect detection of the target nucleic acid utilising the detection probe;
c) incubating the sample solution with the detection probe under conditions for attachment of the detection probe to target nucleic acid in the sample solution;
d) contacting the sample solution with the contact end of the chromatographic strip so that target nucleic acid attached to the detection probe can travel by capillary action to the capture zone and bind to the capture zone by interaction between the hook capture probe immobilised at the capture zone and the first region of the target nucleic acid; and
e) checking for the presence of detection probe at the capture zone.
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12. A method according to claim 12 in which the detection ligand comprises biotin and the detection moiety comprises an anti-biotin antibody, or in which the detection ligand comprises fluorescein and the detection moiety comprises an anti-fluorescein antibody, or in which the detection ligand comprises 2,4-dinitrophenol (DNP) and the detection moiety comprises an anti-DNP antibody.
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18. A method for testing for the presence of target nucleic acid in a sample solution which comprises:
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a) providing a chromatographic strip having a contact end for contacting the sample solution and a capture probe immobilised at a capture zone of the chromatographic strip remote from the contact end, the capture probe being capable of hybridising to a first region of the target nucleic acid;
b) providing a hook detection probe capable of hybridising to a second region of the target nucleic acid;
c) providing a universal detection probe capable of hybridising to the hook detection probe when the hook detection probe has hybridised to the second region of the target nucleic acid, thereby allowing direct or indirect detection of target nucleic acid utilising the universal and hook detection probes;
d) incubating the sample solution with the hook detection probe and the universal detection probe under conditions for hybridisation of the hook detection probe to the second region of the target nucleic acid and for hybridisation of the universal detection probe to the hook detection probe;
e) contacting the sample solution with the contact end of the chromatographic strip so that a complex formed between the universal detection probe, the hook detection probe and target nucleic acid can travel by capillary action to the capture zone and bind to the capture zone by interaction between target nucleic acid of the complex and the capture probe immobilised at the capture zone; and
f) checking for the presence of universal detection probe at the capture zone. - View Dependent Claims (21)
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19. A method for testing for the presence of target nucleic acid in a sample solution which comprises:
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a) providing a chromatographic strip having;
a contact end for contacting the sample solution;
a capture probe immobilised at a capture zone of the chromatographic strip remote from the contact end, the capture probe being capable of hybridising to a first region of the target nucleic acid; and
a universal detection probe releasably immobilised to the chromatographic strip between the contact end and the capture zone;
b) incubating the sample solution with a hook detection probe capable of hybridising to a second region of the target nucleic acid, under conditions for hybridisation of the hook detection probe to the second region;
c) contacting the sample solution with the contact end of the chromatographic strip to cause sample solution to travel by capillary action to the capture zone, thereby releasing universal detection probe so that released universal detection probe can hybridise to hook detection probe in the sample solution as it travels to the capture zone and a complex comprising the universal detection probe, the hook detection probe and target nucleic acid can bind to the capture zone by interaction between target nucleic acid of the complex and the capture probe immobilised at the capture zone; and
f) checking for the presence of universal detection probe at the capture zone.
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20. A method for testing for the presence of target nucleic acid in a sample solution which comprises:
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a) providing a chromatographic strip having a contact end is for contacting the sample solution and a capture probe immobilised at a capture zone of the chromatographic strip remote from the contact end, the capture probe being capable of hybridising to a first region of the target nucleic acid;
b) providing a hook detection probe capable of hybridising to a second region of the target nucleic acid;
c) providing a universal detection probe capable of hybridising to the hook detection probe when the hook detection probe has hybridised to the second region of the target nucleic acid, thereby allowing direct or indirect detection of target nucleic acid utilising the universal and hook detection probes;
d) contacting the sample solution with the contact end of the chromatographic strip to cause sample solution to travel by capillary action to the capture zone and be captured at the capture zone by the capture probe;
e) contacting the capture zone with a hook detection probe and a universal detection probe; and
f) checking for the presence of universal detection probe at the capture zone. - View Dependent Claims (22, 23, 24, 25, 26)
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27. A method for testing for the presence of a target nucleic acid in a sample solution which comprises:
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a) providing a chromatographic strip having a contact end for contacting the sample solution and a capture moiety immobilised at a capture zone of the chromatographic strip remote from the contact end, the capture moiety being capable of binding by non base pairing interaction to a universal capture probe bound to the target nucleic acid by a hook capture probe;
b) providing a hook capture probe capable of hybridising to the target nucleic acid;
c) providing a universal capture probe capable of hybridising to the hook capture probe;
d) contacting the sample solution with;
the hook capture probe under conditions for hybridisation of the hook capture probe to the target nucleic acid; and
the universal capture probe under conditions for hybridisation of the universal capture probe to the hook capture probe;
e) contacting the sample solution with the contact end of the chromatographic strip so that a complex formed between the universal capture probe, the hook capture probe, and target nucleic acid can travel by capillary action to the capture zone and bind to the capture zone by binding (by non base pairing interaction) of the capture moiety to the capture ligand of the complex; and
f) detecting for target nucleic acid at the capture zone.
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28. A dipstick for testing for the presence of target nucleic acid in a sample solution which comprises a chromatographic strip having:
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a contact end for contacting the sample solution;
a universal capture probe immobilised at a capture zone of the chromatographic strip remote from the contact end; and
a hook capture probe hybridised to the universal capture probe and which is capable of hybridising to a first region of the target nucleic acid. - View Dependent Claims (30, 32, 33, 34, 35, 36, 37, 38, 42, 43, 44)
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29. A dipstick for testing for the presence of target nucleic acid in a sample solution which comprises a chromatographic strip having:
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a contact end for contacting the sample solution;
a universal capture probe immobilised at a capture zone of the chromatographic strip remote from the contact end; and
a hook capture probe releasably immobilised to the chromatographic strip between the contact end and the capture zone, the hook capture probe being capable of hybridising to a first region of the target nucleic acid.
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31. A dipstick for testing for the presence of target nucleic acid in a sample solution which comprises a chromatographic strip having:
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a contact end for contacting the sample solution;
a universal capture probe immobilised at a capture zone of the chromatographic strip remote from the contact end, the universal capture probe being capable of hybridising to a hook capture probe bound to a first region of the target nucleic acid; and
a detection moiety releasably immobilised at a conjugate zone of the chromatographic strip between the contact end and the capture zone, the detection moiety being capable of binding by non base pairing interaction to a detection probe hybridised to a second region of the target nucleic acid.
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39. A dipstick for testing for the presence of target nucleic acid in a sample solution which comprises a chromatographic strip having:
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a contact end for contacting the sample solution;
a capture probe immobilised at a capture zone of the chromatographic strip remote from the contact end, the capture probe being capable of binding directly or indirectly to the target nucleic acid;
a universal detection probe releasably immobilised to the chromatographic strip between the contact end and the capture zone, the universal detection probe being capable of hybridising to a hook detection probe; and
optionallya hook detection probe releasably immobilised to the chromatographic strip between the contact end and the capture zone, the hook detection probe being capable of hybridisng to the target nucleic acid. - View Dependent Claims (40)
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41. A dipstick for testing for the presence of a target nucleic acid in a sample solution which comprises:
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a dipstick having a chromatographic strip with a contact end for contacting the sample solution and a capture moiety immobilised at a capture zone of the chromatographic strip remote from the contact end, the capture moiety being 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/or a universal capture probe releasably immobilised to the chromatographic strip between the contact end and the capture zone, the hook capture probe being capable of hybridising to the target nucleic acid, and the universal capture probe being capable of hybridising to the hook capture probe.
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45. A kit for testing for the presence of target nucleic acid in a sample solution which comprises:
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i) a dipstick comprising a chromatographic strip with a contact end for contacting the sample solution, and a universal capture probe immobilised at a capture zone of the chromatographic strip remote from the contact end;
ii) a hook capture probe capable of hybridising to a first region of the target nucleic acid and to the universal capture probe; and
optionallyiii) a detection probe capable of attaching to the target nucleic acid thereby allowing direct or indirect detection of the target nucleic acid utilising the detection probe. - View Dependent Claims (47, 48, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 63)
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46. A kit for testing for the presence of target nucleic acid in a sample solution which comprises:
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i) a dipstick comprising a chromatographic strip with a contact end for contacting the sample solution, a universal capture probe immobilised at a capture zone of the chromatographic strip remote from the contact end, and a hook capture probe hybridised to the universal capture probe, the hook probe being capable of hybridising to a first region of the target nucleic acid; and
optionallyii) a detection probe capable of attaching to the target nucleic acid thereby allowing direct or indirect detection of the target nucleic acid utilising the detection probe.
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49. A kit for testing for the presence of target nucleic acid in a sample solution which comprises:
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i) a dipstick comprising a chromatographic strip with a contact end for contacting the sample solution, and a capture probe immobilised at a capture zone of the chromatographic strip remote from the contact end, the capture probe being capable of hybridising to a first region of the target nucleic acid, or being capable of binding indirectly to the target nucleic acid;
ii) a hook detection probe capable of hybridising to a second region of the target nucleic acid; and
iii) a universal detection probe capable of hybridising to the hook detection probe when the hook detection probe has hybridised to the second region of the target nucleic acid, thereby allowing direct or indirect detection of the target nucleic acid utilising the detection probe.
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60. A kit for testing for the presence of a target nucleic acid in a sample solution which comprises:
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a dipstick having a chromatographic strip with a contact end for contacting the sample solution and a capture moiety immobilised at a capture zone of the chromatographic strip remote from the contact end, the capture moiety being 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 hybridising to the target nucleic acid; and
a universal capture probe capable of hybridising to the hook capture probe.
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61. Use of a universal and a hook capture probe in a dipstick assay for testing for the presence of a target nucleic acid in a sample solution.
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62. Use of a universal and a hook detection probe in a dipstick assay for testing for the presence of a target nucleic acid in a sample solution.
Specification