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EP0387696A2 Expired / Lapsed EP ⏱️ 0.0 years remaining

Method and device for improved reaction kinetics in nucleic acid hybridizations

Assignee: Abbott Laboratories
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Filing: 1990-03-08
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Granted: —
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Expires: 2010-03-08
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Abstract

Abstract The present invention relates to methods and test strips for performing nucleic acid hybridizations on a porous chromatographic material having the capacity for rapid chromatographic solvent transport of non-immobilized reagents and reactive solution components by means of a selected chromatographic solvent. The strip includes a first end or contact site, at which chromatographic solvent transport begins, a second end, at which chromatographic solvent transport ends, and at least one zone positioned between the first and second ends. The zone is impregnated with a capture nucleotide sequence which is immobilized against solvent transport and which is capable of hybridization with a complementary nucleotide sequence or probe so as to render it immobilized in the capture zone. The strip may further include a probe zone between the first end and the capture zone, wherein a labeled probe is impregnated such that it is mobilized by the solvent and transported to the capture zone where hybridization can occur. Interfering sample components and unhybridized nucleic acids are cleared from the capture zone by chromatographic solvent transport to form an inherent wash.

Inventors

  • 👤 Michael Edward Mc Mahon
  • 👤 Julian Gordon

Bibliographic Details

Application Number —
Jurisdiction EP
Data Source google_patents
Last Synchronized 2026-11-05 03:25
CPC Classifications
G; G01; G01N; G01N30/00; C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6876; C12Q1/6888; C12Q1/689; C12Q1/6813; C12Q1/6834; C12Q1/70; C12Q1/701; B; B01; B01L; B01L2400/00; B01L2400/04; B01L2400/0403; B01L2400/0406

Claims Analysis

Total Claims: 12
Claim 1
Independent Claim 87 words
A method of performing hybridization of nucleotide sequences on a solid phase, comprising: a) on a porous medium capable of transporting a solution by capillarity, immobilizing a single stranded nucleotide sequence at a site remote from a contact site of the medium; and b) under hybridizing conditions, contacting the remote site with a solution containing or suspected of containing a complementary nucleotide sequence, the complementary nucleotide sequence being transported by capillarity to the remote site, whereby hybridization can occur between complementary base pairs of the nucleotide sequences.
Claim 2
Independent Claim 87 words
A method of performing hybridization of nucleotide sequences on a solid phase, comprising: a) on a porous medium capable of transporting a solution by capillarity, immobilizing a single stranded nucleotide sequence at a site remote from a contact site of the medium; and b) under hybridizing conditions, contacting the remote site with a solution containing or suspected of containing a complementary nucleotide sequence, the complementary nucleotide sequence being transported by capillarity to the remote site, whereby hybridization can occur between complementary base pairs of the nucleotide sequences.
Claim 3
Dependent Claim 36 words
A method according to claim 1, wherein the solution containing or suspected of containing complementary nucleotide sequence is contacted with the medium at the contact site and is transport to the remote site in the solution.
Claim 4
Dependent Claim 46 words
A method according to claim 1, comprising an additional step of drying the complementary nucleotide sequence onto the medium at a location between the contact site and the remote site, so that the complementary nucleotide sequence is mobilized and transported to the remote site in solution.
Claim 5
Dependent Claim 31 words
A method according to claim 3, comprising a step of contacting the contact site with a solvent capable of mobilizing the complementary nucleotide sequence and transporting it to the remote site.
Claim 6
Dependent Claim 16 words
A method according to claim 1 and comprising a step of detecting the complementary nucleotide sequence.
Claim 7
Dependent Claim 37 words
A method according to claim 1 wherein the solution containing the complementary nucleotide sequence is transported through the medium at a rate such that a solution front contacts the remote site for less than about 10 minutes.
Claim 8
Dependent Claim 37 words
A method according to claim 1 wherein the solution containing the complementary nucleotide sequence is transported through the medium at a rate such that a solution front contacts the remote site for less than about 2 minutes.
Claim 9
Independent Claim 75 words
A device for performing solid phase hybridizations of soluble, single stranded nucleotide sequences, comprising a porous medium having: a contact site; a site remote from the contact site whereat a single stranded nucleotide sequence complementary to a predetermined soluble nucleotide sequence is immobilized; and capillary means for transporting a solution containing the predetermined soluble nucleotide sequence under hybridizing conditions to the remote site whereby hybridization can occur between complementary base pairs of the nucleotide sequences.
Claim 10
Independent Claim 75 words
A device for performing solid phase hybridizations of soluble, single stranded nucleotide sequences, comprising a porous medium having: a contact site; a site remote from the contact site whereat a single stranded nucleotide sequence complementary to a predetermined soluble nucleotide sequence is immobilized; and capillary means for transporting a solution containing the predetermined soluble nucleotide sequence under hybridizing conditions to the remote site whereby hybridization can occur between complementary base pairs of the nucleotide sequences.
Claim 11
Dependent Claim 43 words
A device according to claim 8, further comprising soluble nucleotide sequence dried on the medium at a location between the contact site and the remote site, such that it can be mobilized by the solution and transported to the remote site by capillarity.
Claim 12
Dependent Claim 23 words
A device according to claim 8 wherein the porous medium comprises a strip of nitrocellulose having a pore size of about 5.0 microns.

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