Hybridization method for the detection of genetic material

Claim 1

A method for the detection of single-stranded target genetic material comprising the steps of: (a) contacting, under conditions permissive of hybridization, said target genetic material with (i) a first polynucleotide probe comprising a first partic to which is attached a plurality of first single-stranded polynucleotide segments complementary to a first portion of said target genetic material, and (ii) a second polynucleotide probe comprising a second particle having attached thereto a plurality of second single-stranded polynucleotide segments complementary to a second portion of said target genetic material; (b) permitting hybridization of said first polynucleotide segments to said first portion of target genetic material and said second segments to said second portion of target genetic material, said hybridizations resulting in agglutination; and (c) detecting said agglutination.

Claim 2

A method for the detection of single-stranded target genetic material comprising the steps of: (a) contacting, under conditions permissive of hybridization, said target genetic material with (i) a first polynucleotide probe comprising a first partic to which is attached a plurality of first single-stranded polynucleotide segments complementary to a first portion of said target genetic material, and (ii) a second polynucleotide probe comprising a second particle having attached thereto a plurality of second single-stranded polynucleotide segments complementary to a second portion of said target genetic material; (b) permitting hybridization of said first polynucleotide segments to said first portion of target genetic material and said second segments to said second portion of target genetic material, said hybridizations resulting in agglutination; and (c) detecting said agglutination.

Claim 6

A method for the detection of single-stranded target genetic material comprising the steps of: (a) contacting, under conditions permissive of hybridization. (i) an unhybridized portion of a first strand of target genetic material with a first polynucleotide probe comprising a first particle to which is attached a plurality of first single-stranded polynucleotide segments complementary to said first single strand of target genetic material; (ii) an unhybridized portion of a second strand of target genetic material with a second polynucleotide probe comprising a second particle having attached thereto a plurality of second single-stranded polynucleotide segments complementary to said second single strand of target genetic material; and (iii) an unhybridized portion of said first and said second strand of target genetic material; with the provisos that: said first and second single stranded polynucleotide segments are not capable of hybridizing to each other: hybridization of said first polynucleotide segment to said unhybridized portion of said first strand of target genetic material and hybridization of said second polynucleotide segment to said unhybridized portion of said second strand of target genetic material do not prevent at least a portion of said first strand of target genetic material from being available to hybridize with at least a portion of said second strand of target genetic material, (b) permitting the hybridization of (1) said first polynucleotide segment to said unhybridized portion of said first strand of target genetic material, (2) said second polynucleotide segment to said unhybridized portion of said second strand of target genetic material, and (3) said unhybridized portion of said first strand of target genetic material and said unhybridized portion of said second strand of target genetic material, the foregoing hybridizations occurring in any order; said hybridizations resulting in agglutination; and (c) detecting such hybridizations by means of the agglutination that occurs.

Claim 7

A method for the detection of single-stranded target genetic material comprising the steps of: (a) contacting, under conditions permissive of hybridization. (i) an unhybridized portion of a first strand of target genetic material with a first polynucleotide probe comprising a first particle to which is attached a plurality of first single-stranded polynucleotide segments complementary to said first single strand of target genetic material; (ii) an unhybridized portion of a second strand of target genetic material with a second polynucleotide probe comprising a second particle having attached thereto a plurality of second single-stranded polynucleotide segments complementary to said second single strand of target genetic material; and (iii) an unhybridized portion of said first and said second strand of target genetic material; with the provisos that: said first and second single stranded polynucleotide segments are not capable of hybridizing to each other: hybridization of said first polynucleotide segment to said unhybridized portion of said first strand of target genetic material and hybridization of said second polynucleotide segment to said unhybridized portion of said second strand of target genetic material do not prevent at least a portion of said first strand of target genetic material from being available to hybridize with at least a portion of said second strand of target genetic material, (b) permitting the hybridization of (1) said first polynucleotide segment to said unhybridized portion of said first strand of target genetic material, (2) said second polynucleotide segment to said unhybridized portion of said second strand of target genetic material, and (3) said unhybridized portion of said first strand of target genetic material and said unhybridized portion of said second strand of target genetic material, the foregoing hybridizations occurring in any order; said hybridizations resulting in agglutination; and (c) detecting such hybridizations by means of the agglutination that occurs.

Claim 11

A method for the detection of single-stranded target genetic material which comprises: (a) Contacting complementary strands of said target genetic material under conditions permissive of hybridization, with an amount, relative to the amount of target genetic material sufficient to prevent substantial reannealing thereof, of a polynucleotide probe wherein said polynucleotide probe comprises a label attached to a single-stranded polynucleotide segment having a first portion complementary to a first strand of the target genetic material and a second portion complementary to a second strand of the target genetic material, said first strand and said second strand of target genetic material being (I) complementary to each other and (ii) such that when said first portion and said second portion are hybridized to said target genetic material, there is at least one sequence of each of the complementary strands of said target genetic material that is available to hybridize to the other strand; (b) permitting hybridization thereof resulting in the formation of a detectable multihybrid, and (c) detecting said multihybrid.

Claim 12

A method for the detection of single-stranded target genetic material which comprises: (a) Contacting complementary strands of said target genetic material under conditions permissive of hybridization, with an amount, relative to the amount of target genetic material sufficient to prevent substantial reannealing thereof, of a polynucleotide probe wherein said polynucleotide probe comprises a label attached to a single-stranded polynucleotide segment having a first portion complementary to a first strand of the target genetic material and a second portion complementary to a second strand of the target genetic material, said first strand and said second strand of target genetic material being (I) complementary to each other and (ii) such that when said first portion and said second portion are hybridized to said target genetic material, there is at least one sequence of each of the complementary strands of said target genetic material that is available to hybridize to the other strand; (b) permitting hybridization thereof resulting in the formation of a detectable multihybrid, and (c) detecting said multihybrid.

Claim 18

A method for the detection of a single-stranded target genetic material which comprises (a) contacting, under conditions permissive of hybridization, a sample suspected of containing said target genetic material with (i) a plurality of first polynucleotide probes comprising a first entity attached to a first single-stranded polynucleotide segment which is hybridizable with a first portion of said target genetic material; (ii) a plurality of first particles having attached thereto a second entity which is specifically bindable with said first entity; (iii) a plurality of second polynucleotide probes comprising a third entity and attached thereto a second single-stranded polynucleotide segment hybridizable with a second portion of said target genetic material; and (iv) a plurality of second particles having attached thereto a fourth entity which is specifically bindable with said third entity; (b) permitting hybridization of said first and second probes with said target genetic material, binding of said first entity with said second entity and binding of said third entity with said fourth entity so as to form an agglutination complex; and (c) detecting the presence of said target by observing said agglutination complex.

Claim 19

A method for the detection of a single-stranded target genetic material which comprises (a) contacting, under conditions permissive of hybridization, a sample suspected of containing said target genetic material with (i) a plurality of first polynucleotide probes comprising a first entity attached to a first single-stranded polynucleotide segment which is hybridizable with a first portion of said target genetic material; (ii) a plurality of first particles having attached thereto a second entity which is specifically bindable with said first entity; (iii) a plurality of second polynucleotide probes comprising a third entity and attached thereto a second single-stranded polynucleotide segment hybridizable with a second portion of said target genetic material; and (iv) a plurality of second particles having attached thereto a fourth entity which is specifically bindable with said third entity; (b) permitting hybridization of said first and second probes with said target genetic material, binding of said first entity with said second entity and binding of said third entity with said fourth entity so as to form an agglutination complex; and (c) detecting the presence of said target by observing said agglutination complex.