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EP0159719A2 Expired / Lapsed EP ⏱️ 0.0 years remaining

Hybridization method for the detection of genetic materials

Assignee: Enzo Biochem Inc
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Filing: 1985-04-26
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Granted: —
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Expires: 2005-04-26
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Abstract

Abstract This invention provides methods for the detection of a target genetic material having a desired base sequence or gene. Also disclosed are methods for the detection of mutations. Also provided are components for use in such methods. The methods are based upon techniques which utilize two labeled single stranded polynucleotide segments which are complementary to the same or the opposite strands of the target genetic material. The methods of the invention result in the formation of a double hybrid and/or a multihybrid.

Inventors

  • 👤 Elazar Rabbani
  • 👤 Dean L. Engelhardt

Bibliographic Details

Application Number —
Jurisdiction EP
Data Source google_patents
Last Synchronized 2026-11-06 13:40
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6813; C12Q1/6816; C12Q1/682; C12Q1/6827; C12Q1/683; C12Q1/6832; Y; Y10; Y10S; Y10S435/00; Y10S435/803

Claims Analysis

Total Claims: 17
Claim 1
Independent Claim 134 words
A method for the detection of target genetic material which comprises: providing (A) a first polynucleotide probe comprising at least one first label and at least one first single stranded polynucleotide segment attached to said first label, and (B) a second polynucleotide probe comprising at least one second label and at least one second single stranded polynucleotide segment attached to said second label, wherein said first segment and said second segment are complementary to substantially mutually exclusive portions of the same strand of said target material; rendering said target material single stranded; contacting said first probe and said second probe with said single-stranded target material under hybridizing conditions to form a double hybrid or a multihybrid; and detecting said double hybrid or said multihybrid by means of said first label and said second label.
Claim 2
Independent Claim 134 words
A method for the detection of target genetic material which comprises: providing (A) a first polynucleotide probe comprising at least one first label and at least one first single stranded polynucleotide segment attached to said first label, and (B) a second polynucleotide probe comprising at least one second label and at least one second single stranded polynucleotide segment attached to said second label, wherein said first segment and said second segment are complementary to substantially mutually exclusive portions of the same strand of said target material; rendering said target material single stranded; contacting said first probe and said second probe with said single-stranded target material under hybridizing conditions to form a double hybrid or a multihybrid; and detecting said double hybrid or said multihybrid by means of said first label and said second label.
Claim 3
Dependent Claim 102 words
The method according to Claim 1, which further comprises: providing (A) a third polynucleotide probe comprising at least one third label and at least one third single stranded polynucleotide segment attached to said third label, wherein said third segment is complementary to said first segment, and (B) a'fourth polynucleotide probe comprising at least one fourth label and at least one fourth single stranded polynucleotide segment attached to said fourth label, wherein said fourth segment is complementary to said second segment; wherein said first label and said third label comprise chemoluminescent catalysts and said second label and said fourth label comprise absorber/emitter moieties.
Claim 4
Independent Claim 166 words
A method for the detection of target genetic material which comprises: providing a first polynucleotide probe comprising at least one first label and at least one first single stranded polynucleotide segment attached to said first label, said first segment being complementary to a first portion of one strand of said target material; providing a second polynucleotide probe comprising at least one second label and at least one second single stranded polynucleotide segment attached to said second label, said second segment being complementary to a second portion of the opposite strand of said target material; said first segment being non-complementary to said second segment and said second portion, and said second segment being non-complementary to said first portion; rendering said target material single stranded; contacting said first probe and said second probe with said single stranded target material under hybridizing conditions to form a double hybrid or a multihybrid; and detecting said double hybrid or said multihybrid by means of said first label and said second label.
Claim 5
Independent Claim 166 words
A method for the detection of target genetic material which comprises: providing a first polynucleotide probe comprising at least one first label and at least one first single stranded polynucleotide segment attached to said first label, said first segment being complementary to a first portion of one strand of said target material; providing a second polynucleotide probe comprising at least one second label and at least one second single stranded polynucleotide segment attached to said second label, said second segment being complementary to a second portion of the opposite strand of said target material; said first segment being non-complementary to said second segment and said second portion, and said second segment being non-complementary to said first portion; rendering said target material single stranded; contacting said first probe and said second probe with said single stranded target material under hybridizing conditions to form a double hybrid or a multihybrid; and detecting said double hybrid or said multihybrid by means of said first label and said second label.
Claim 6
Dependent Claim 46 words
The method according to Claim 1 or 3, characterized in that said first label is a microparticle, said second label is a moiety that is capable of creating a signal, and said detecting step further comprises separating unhybridized second probes from said double hybrids or multihybrids.
Claim 7
Independent Claim 107 words
A method for the detection of target genetic material which comprises: rendering said target material single stranded; contacting said single stranded target material with a polynucleotide probe under hybridizing conditions, said probe comprising at least one first label and at least one single stranded polynucleotide segment attached to said first label, said segment being complementary to a portion of one strand of said target material, thereby forming double stranded genetic material; contacting said double stranded material with an antibody directed to double stranded genetic material under conditions permitting said antibody to bind to said double stranded material, thereby forming a bound entity; and detecting said bound entity.
Claim 8
Independent Claim 107 words
A method for the detection of target genetic material which comprises: rendering said target material single stranded; contacting said single stranded target material with a polynucleotide probe under hybridizing conditions, said probe comprising at least one first label and at least one single stranded polynucleotide segment attached to said first label, said segment being complementary to a portion of one strand of said target material, thereby forming double stranded genetic material; contacting said double stranded material with an antibody directed to double stranded genetic material under conditions permitting said antibody to bind to said double stranded material, thereby forming a bound entity; and detecting said bound entity.
Claim 9
Independent Claim 66 words
A method for the detection of target genetic material which comprises: providing a polynucleotide probe comprising a single stranded polynucleotide segment having noncontiguous first and second portions complementary to substantially mutually exclusive portions of a single strand of said target material; rendering said target material single stranded; contacting said probe with said single-stranded target material under hybridizing conditions to form a multihybrid, and detecting said multihybrid.
Claim 10
Independent Claim 66 words
A method for the detection of target genetic material which comprises: providing a polynucleotide probe comprising a single stranded polynucleotide segment having noncontiguous first and second portions complementary to substantially mutually exclusive portions of a single strand of said target material; rendering said target material single stranded; contacting said probe with said single-stranded target material under hybridizing conditions to form a multihybrid, and detecting said multihybrid.
Claim 11
Independent Claim 125 words
A method for the detection of target genetic material which comprises: providing a polynucleotide probe comprising a single stranded polynucleotide segment comprising a first segment portion complementary to a first portion of a strand of said target material and a second segment portion complementary to a second portion of the opposite strand of said target material, wherein when said first and second segment portions are hybridized to said first and second strand portions, at least one single stranded portion present on said first strand is capable of hybridizing to at least one complementary single-stranded portion present on said opposite strand; rendering said target material single stranded; contacting said probe with said single-stranded target material under hybridizing conditions to form a multihybrid; and detecting said multihybrid.
Claim 12
Independent Claim 125 words
A method for the detection of target genetic material which comprises: providing a polynucleotide probe comprising a single stranded polynucleotide segment comprising a first segment portion complementary to a first portion of a strand of said target material and a second segment portion complementary to a second portion of the opposite strand of said target material, wherein when said first and second segment portions are hybridized to said first and second strand portions, at least one single stranded portion present on said first strand is capable of hybridizing to at least one complementary single-stranded portion present on said opposite strand; rendering said target material single stranded; contacting said probe with said single-stranded target material under hybridizing conditions to form a multihybrid; and detecting said multihybrid.
Claim 13
Independent Claim 126 words
A method for the detection of target genetic material which comprises: providing (A) a first polynucleotide probe comprising at least one label, and at least one first single stranded polynucleotide segment attached to said label, and (B) a second polynucleotide probe comprising at least one second single stranded polynucleotide segment having attached thereto a first moiety that is capable of forming a complex with a second moiety; wherein said first and second segments are complementary to substantially mutually exclusive portions of a single strand of said target material or said first segment is complementary to a first portion of said strand and said second segment is complementary to a second portion of the opposite strand, said first strand portion being non-complementary to said second strand portion.
Claim 14
Independent Claim 126 words
A method for the detection of target genetic material which comprises: providing (A) a first polynucleotide probe comprising at least one label, and at least one first single stranded polynucleotide segment attached to said label, and (B) a second polynucleotide probe comprising at least one second single stranded polynucleotide segment having attached thereto a first moiety that is capable of forming a complex with a second moiety; wherein said first and second segments are complementary to substantially mutually exclusive portions of a single strand of said target material or said first segment is complementary to a first portion of said strand and said second segment is complementary to a second portion of the opposite strand, said first strand portion being non-complementary to said second strand portion.
Claim 15
Dependent Claim 100 words
A method according to claim 8, characterized in that said first and second segments are complementary to portions of a single strand of said target material, and further comprising the steps of: providing a matrix which has said second moiety attached thereto; rendering said target material single stranded; forming an entity comprising (i) said first probe hybridized to a single strand of said target material through said first segment, (ii) said second probe hybridized to said strand through said second segment; and (iii) said first moiety complexed through said second moiety; and detecting said entity by means of said label.
Claim 16
Dependent Claim 50 words
A method according to Claim 8 or 9, characterized in that said first probe is hybridized to said target material through said first segment and said second probe is hybridized to said target material through said second segment prior to the complexation of said first moiety and said second moiety.
Claim 17
Independent Claim 37 words
The method according to any of claims 1 to 10, characterized in that said target genetic material is selected from the group consisting of double-stranded or partially double-stranded DNA, RNA, oligo- or polynucleotides, gene sequences and proteins.

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