EP0128018A2
Expired / Lapsed
EP
⏱️ 0.0 years remaining
Molecular genetic probe, and method of forming same, assay technique and kit using said molecular genetic probe
Assignee: Orgenics Ltd
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Filing: 1984-05-31
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Granted: —
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Expires: 2004-05-31
Abstract
Abstract A method of assaying for the presence or quantity of a suspect nucleic acid in a suspect sample using a modified genetic probe in combination with an imaging procedure. The probe is modified by addition and provides an easy and very effective technique of modifying a genetic probe preparatory to its use in a kit. The genetic probe is then introduced into contact with DNA or RNAofthesuspectsample and permitted to hybridize therewith. A kit for performing an assay using the modified probe is also disclosed.
Inventors
- 👤 Max Herzberg
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-28 20:31
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6804; C12Q1/6813; C12Q1/6816
Claims Analysis
Total Claims: 58
Claim 1
Independent Claim
135 words
1) A method of assay for the presence or quantity of a suspect nucleic acid in a suspect sample in which the suspect sample is contacted with a modified nucleic acid selected such that it will hybridize with the suspect nucleic acid, if it is present, to form a hybridized complex, characterised in that, a) the modified nucleic acid has been formed by a one-step modification of at least one base of the nucleic acid; b) the modified nucleic acid is contacted with labelled antibodies selected to complex with at least the modified portion of the modified nucleic acid to form a hybridized complex; and c) the presence and/or extent of the presence of the labelled antibodies is measured to obtain an indication of the presence, or extent of presence, of the suspect nucleic acid.
Claim 2
Dependent Claim
135 words
1) A method of assay for the presence or quantity of a suspect nucleic acid in a suspect sample in which the suspect sample is contacted with a modified nucleic acid selected such that it will hybridize with the suspect nucleic acid, if it is present, to form a hybridized complex, characterised in that, a) the modified nucleic acid has been formed by a one-step modification of at least one base of the nucleic acid; b) the modified nucleic acid is contacted with labelled antibodies selected to complex with at least the modified portion of the modified nucleic acid to form a hybridized complex; and c) the presence and/or extent of the presence of the labelled antibodies is measured to obtain an indication of the presence, or extent of presence, of the suspect nucleic acid.
Claim 3
Dependent Claim
27 words
The method of claim 1 comprising contacting said modified nucleic acid with said labelled antibodies after said modified nucleic acid and said suspect sample have been contacted.
Claim 4
Dependent Claim
140 words
3) A method of assaying for the presence or quantity of a suspect nucleic acid in which the suspect sample is contacted with a modified nucleic acid selected such that it will hybridize with said suspect nucleic acid, if it is present, to form a hybridized complex, characterised in that,: a) the modified nucleic acid has been formed by a one-step modification of at least one base of said nucleic acid; b) the modified nucleic acid is contacted with an identifying agent selected to complex with at least the modified portion of the modified nucleic acid, said identifying agent being selected to signal the presence of said hybridized complex; and c) the presence and/or extent of the presence of said identifying agent is measured to obtain an indication of the presence or extent of presence of said suspect nucleic acid.
Claim 5
Dependent Claim
140 words
3) A method of assaying for the presence or quantity of a suspect nucleic acid in which the suspect sample is contacted with a modified nucleic acid selected such that it will hybridize with said suspect nucleic acid, if it is present, to form a hybridized complex, characterised in that,: a) the modified nucleic acid has been formed by a one-step modification of at least one base of said nucleic acid; b) the modified nucleic acid is contacted with an identifying agent selected to complex with at least the modified portion of the modified nucleic acid, said identifying agent being selected to signal the presence of said hybridized complex; and c) the presence and/or extent of the presence of said identifying agent is measured to obtain an indication of the presence or extent of presence of said suspect nucleic acid.
Claim 6
Dependent Claim
31 words
4) The method of claim 3 comprising contacting said modified nucleic acid with said suspect sample to form said hybridized complex prior to contacting said modified nucleic acid with said antibodies.
Claim 7
Dependent Claim
127 words
5) A method of assaying for the presence or quantity of a suspect nucleic acid in which the suspect sample is contacted with a modified nucleic acid selected such that it will hybridize with said suspect nucleic acid, if it is present, to form a hybridized complex, characterised in that,: a) the modified nucleic acid has been formed by a non-enzymatic modification of at least one base of said nucleic acid; b) the modified nucleic acid is contacted with a labelling agent selected to complex with at least the modified portion of said modified nucleic acid; and c) the presence and/or extent of the presence of said labelling agent is measured to obtain an indication of the presence or extent of presence of said suspect nucleic acid.
Claim 8
Dependent Claim
127 words
5) A method of assaying for the presence or quantity of a suspect nucleic acid in which the suspect sample is contacted with a modified nucleic acid selected such that it will hybridize with said suspect nucleic acid, if it is present, to form a hybridized complex, characterised in that,: a) the modified nucleic acid has been formed by a non-enzymatic modification of at least one base of said nucleic acid; b) the modified nucleic acid is contacted with a labelling agent selected to complex with at least the modified portion of said modified nucleic acid; and c) the presence and/or extent of the presence of said labelling agent is measured to obtain an indication of the presence or extent of presence of said suspect nucleic acid.
Claim 9
Dependent Claim
32 words
6) The method of claim 5 comprising contacting said modified nucleic acid with said suspect sample to form said hybridized complex prior to contacting said modified nucleic acid with said labelling agent
Claim 10
Dependent Claim
121 words
7) A method of assaying for the presence or quantity of a suspect nucleic acid in which the suspect sample is contacted with a modified nucleic acid selected such that it will hybridize with said suspect nucleic acid, if it is present, to form a hybridized complex, characterised in that,: a) the modified nucleic acid has a modifying agent coupled thereto without a linker arm; b) the modified nucleic acid is contacted with a labelling agent selected to complex with at least the modified portion of said modified nucleic acid; and c) the presence and/or extent of the presence of said labelling agent is measured to obtain an indication of the presence or extent of presence of said suspect nucleic acid.
Claim 11
Dependent Claim
121 words
7) A method of assaying for the presence or quantity of a suspect nucleic acid in which the suspect sample is contacted with a modified nucleic acid selected such that it will hybridize with said suspect nucleic acid, if it is present, to form a hybridized complex, characterised in that,: a) the modified nucleic acid has a modifying agent coupled thereto without a linker arm; b) the modified nucleic acid is contacted with a labelling agent selected to complex with at least the modified portion of said modified nucleic acid; and c) the presence and/or extent of the presence of said labelling agent is measured to obtain an indication of the presence or extent of presence of said suspect nucleic acid.
Claim 12
Dependent Claim
32 words
8) The method of claim 7, comprising contacting said modified nucleic acid with said suspect sample to form said hybridized complex prior to contacting said modified nucleic acid with said labelling agent.
Claim 13
Dependent Claim
18 words
9) The method of any of claims 1-8, wherein said modified nucleic acid has at least 10 bases.
Claim 14
Dependent Claim
18 words
10) The method of any of claims 1-9, wherein said modified nucleic acid has at least 50 bases.
Claim 15
Dependent Claim
18 words
11) The method of any of claims 1-10, wherein said suspect nucleic acid has at least 10 bases.
Claim 16
Dependent Claim
18 words
12) The method of any of claims 1-11, wherein said suspect nucleic acid codes for a gene product.
Claim 17
Dependent Claim
17 words
13) The method of any of claims 1-12, wherein said suspect nucleic acid codes for an enzyme.
Claim 18
Dependent Claim
16 words
14) The method of any of claims 1-13, wherein said suspect nucleic acid is viral DNA.
Claim 19
Dependent Claim
15 words
15) The method of claim 14, wherein said suspect nucleic acid is viral Herpes DNA.
Claim 20
Dependent Claim
18 words
16) The method of any of claims 1-15, wherein said modified nucleic acid has been modified by alkylation.
Claim 21
Dependent Claim
27 words
17) The method of claim 16, wherein said modified nucleic acid has been modified by alkylation of the 0 6 position of at least one guanidine residue.
Claim 22
Dependent Claim
13 words
18) The method of claim 16 wherein said modified nucleic acid is butylated.
Claim 23
Dependent Claim
13 words
19) The method of claim 16 wherein said modified nucleic acid is ethylated.
Claim 24
Dependent Claim
19 words
20) The method of any one of claims 1-15, wherein said modified nucleic acid has been modified by sulfonation.
Claim 25
Dependent Claim
19 words
21) The method of any one of claims 1-15, wherein said modified nucleic acid has been modified by nitrosonation.
Claim 26
Dependent Claim
19 words
22) The method of any one of claims 1-15, wherein said modified nucleic acid has been formed by nitrophenylation.
Claim 27
Dependent Claim
20 words
23) The method of any one of claims 1-15, wherein said modified nucleic acid was formed by an addition reaction.
Claim 28
Dependent Claim
25 words
24) The method of claim 1 comprising performing step b) wherein said labelled antibodies are labelled with an enzyme selected to provide a color reaction.
Claim 29
Dependent Claim
22 words
25) The method of claim 24 wherein said enzyme is selected from the group consisting of peroxidase, beta-galactosidase, hyaluronidase, or alkaline phosphatase
Claim 30
Dependent Claim
18 words
26) The method of any one of claims 1-25, wherein said suspect nucleic acid is single stranded RNA.
Claim 31
Dependent Claim
36 words
27) The method of any one of claims 1-25, wherein said suspect nucleic acid is normally double stranded and was at least partially denatured prior to contact with said modified nucleic acid according to step a).
Claim 32
Dependent Claim
146 words
28) A method of assaying for the presence or quantity of a suspect nucleic acid in a suspect sample characterised in that it comprises the steps of;- a) modifying a nucleic acid capable of hybridizing with said suspect nucleic acid to form a modified nucleic acid substantially without the use of mercury; b) contacting said suspect sample with said modified nucleic acid to form a hybridized complex; c) contacting said modified nucleic acid with antibodies selected to complex with at least the modified portion of said modified nucleic acid, whereby at least one of said antibodies may complex with said modified nucleic acid; d) labelling said antibodies with an agent adapted to signal the presence of said antibodies; and e) measuring for the presence and/or extent of thepresence of said label as an indication of the presence of extent of presence of said suspect nucleic acid.
Claim 33
Dependent Claim
146 words
28) A method of assaying for the presence or quantity of a suspect nucleic acid in a suspect sample characterised in that it comprises the steps of;- a) modifying a nucleic acid capable of hybridizing with said suspect nucleic acid to form a modified nucleic acid substantially without the use of mercury; b) contacting said suspect sample with said modified nucleic acid to form a hybridized complex; c) contacting said modified nucleic acid with antibodies selected to complex with at least the modified portion of said modified nucleic acid, whereby at least one of said antibodies may complex with said modified nucleic acid; d) labelling said antibodies with an agent adapted to signal the presence of said antibodies; and e) measuring for the presence and/or extent of thepresence of said label as an indication of the presence of extent of presence of said suspect nucleic acid.
Claim 34
Dependent Claim
30 words
29) The method of claim 28 comprising contacting said modified nucleic acid and said suspect sample prior to contacting said modified nucleic acid with said antibodies, according to step c).
Claim 35
Dependent Claim
13 words
30) The method of claims 28-29 comprising modifying said nucleic acid by alkylation.
Claim 36
Dependent Claim
23 words
31) The method of claim 30 comprising alkylating said nucleic acid with a compound selected from the group consisting of methyl-,ethyl- or propylnitrosourea.
Claim 37
Dependent Claim
124 words
32) A method of assaying for the presence or quantity of a suspect nucleic acid in a suspect sample characterised in that it comprises the steps of:- a) modifying a double-stranded hybridizable nucleic acid which is hybridizable with said suspect nucleic acid by non-enzymatically adding a moiety from a base of sdaid nucleic acid to form a modified nucleic acid; b) contacting said suspect sample with said modified nucleic acid; c) contacting said modified nucleic acid with a labelling agent selected to complex with at least the modified portion of said modified nucleic acid; and d) measuring for the presence and/or extent of the presence of said labelling agent as an indication of the presence or extent of presence of said suspect nucleic acid.
Claim 38
Dependent Claim
124 words
32) A method of assaying for the presence or quantity of a suspect nucleic acid in a suspect sample characterised in that it comprises the steps of:- a) modifying a double-stranded hybridizable nucleic acid which is hybridizable with said suspect nucleic acid by non-enzymatically adding a moiety from a base of sdaid nucleic acid to form a modified nucleic acid; b) contacting said suspect sample with said modified nucleic acid; c) contacting said modified nucleic acid with a labelling agent selected to complex with at least the modified portion of said modified nucleic acid; and d) measuring for the presence and/or extent of the presence of said labelling agent as an indication of the presence or extent of presence of said suspect nucleic acid.
Claim 39
Dependent Claim
32 words
33) The method of claim 32 comprising contacting said modified nucleic acid with said suspect ample to form said hybridized complex prior to contacting said modified nucleic acid with said labelling agent.
Claim 40
Dependent Claim
161 words
34) A kit for testing for the presence of a nucleic acid in a suspect sample characterised in that it comprises:- a) a supply of modified nucleic acid selected so that it will hybridize with said suspect nucleic acid, it it is present, to form a hybridized complex, said modified nucleic acid having been formed by a one-step modification of a base of said nucleic acid; b) means for contacting said suspect sample with said modified nucleic acid to form said hybridized complex: c) a supply of antibodies selected to complex with at least the modified portion of said modified nucleic acid, said antibodies being labelled with a labelling agent adapted to signal the presence of said antibody; d) means for contacting said modified nucleic acid with said labelled antibodies; and e) means for measuring for the presence and/or extent of the presence of said label as an indication of the presence or extent of presence of said suspect nucleic acid.
Claim 41
Dependent Claim
161 words
34) A kit for testing for the presence of a nucleic acid in a suspect sample characterised in that it comprises:- a) a supply of modified nucleic acid selected so that it will hybridize with said suspect nucleic acid, it it is present, to form a hybridized complex, said modified nucleic acid having been formed by a one-step modification of a base of said nucleic acid; b) means for contacting said suspect sample with said modified nucleic acid to form said hybridized complex: c) a supply of antibodies selected to complex with at least the modified portion of said modified nucleic acid, said antibodies being labelled with a labelling agent adapted to signal the presence of said antibody; d) means for contacting said modified nucleic acid with said labelled antibodies; and e) means for measuring for the presence and/or extent of the presence of said label as an indication of the presence or extent of presence of said suspect nucleic acid.
Claim 42
Dependent Claim
46 words
35) A single stranded DNA molecular genetic probe characterised in that it comprises at least one modified base, said base being modified by a one-step addition while it is attached to said DNA, said molecular genetic probe being adapted to hybridize with a complementary DNA strand.
Claim 43
Dependent Claim
46 words
35) A single stranded DNA molecular genetic probe characterised in that it comprises at least one modified base, said base being modified by a one-step addition while it is attached to said DNA, said molecular genetic probe being adapted to hybridize with a complementary DNA strand.
Claim 44
Dependent Claim
16 words
36) A labelled antibody specifically formed to complex with the molecular genetic probe of claim 35.
Claim 45
Dependent Claim
48 words
37) A single stranded RNA molecular genetic probe characterised in that it comprises at least one modified base, said base being modified by a one-step addition while it is attached to said RNA, said molecular genetic probe being adapted to hybridize with a complementary DNA or RNA strand.
Claim 46
Dependent Claim
48 words
37) A single stranded RNA molecular genetic probe characterised in that it comprises at least one modified base, said base being modified by a one-step addition while it is attached to said RNA, said molecular genetic probe being adapted to hybridize with a complementary DNA or RNA strand.
Claim 47
Dependent Claim
16 words
38) A labelled antibody specifically formed to complex with the molecular genetic probe of claim 37.
Claim 48
Dependent Claim
43 words
39) A method of forming a modified molecular genetic probe characterised in that it comprises the steps of adding a moiety onto a base of a polynucleotide in a single step to form a modified polynucleotide capable of hybridizing with a suspect polynucleotide.
Claim 49
Dependent Claim
43 words
39) A method of forming a modified molecular genetic probe characterised in that it comprises the steps of adding a moiety onto a base of a polynucleotide in a single step to form a modified polynucleotide capable of hybridizing with a suspect polynucleotide.
Claim 50
Dependent Claim
45 words
40') A method of forming a modified molecular genetic probe characterised in that it comprises the step of adding a moiety onto a base of a polynucleotide substantially without the use of mercury to form a modified polynucleotide capable of hybridizing with a suspect polynucleotide.
Claim 51
Dependent Claim
45 words
40') A method of forming a modified molecular genetic probe characterised in that it comprises the step of adding a moiety onto a base of a polynucleotide substantially without the use of mercury to form a modified polynucleotide capable of hybridizing with a suspect polynucleotide.
Claim 52
Dependent Claim
53 words
41) A method of forming a modified molecular genetic probe characterised in that it comprises the step of adding a moiety onto a base of a double stranded polynucleotide non-enzymatically to form a modified double stranded polynucleotide and at least partially denaturing said modified double stranded polynucleotide to form said modified genetic probe
Claim 53
Dependent Claim
53 words
41) A method of forming a modified molecular genetic probe characterised in that it comprises the step of adding a moiety onto a base of a double stranded polynucleotide non-enzymatically to form a modified double stranded polynucleotide and at least partially denaturing said modified double stranded polynucleotide to form said modified genetic probe
Claim 54
Dependent Claim
44 words
42) A single stranded molecular genetic probe characterised in that it comprises at least one base modified by a moiety, said moiety being added directly to said base without a linker arm, said molecular genetic probe being adapted to hybridize with a complementary polynucleotide.
Claim 55
Dependent Claim
44 words
42) A single stranded molecular genetic probe characterised in that it comprises at least one base modified by a moiety, said moiety being added directly to said base without a linker arm, said molecular genetic probe being adapted to hybridize with a complementary polynucleotide.
Claim 56
Dependent Claim
141 words
43) A method of assaying for the presence or quantity of a suspect nucleic acid in a suspect sample characterised in that it comprises the steps of: a) contacting said suspect sample with a plurality of modified nucleic acids, each of said nucleic acids being selected such that it may hybridize with said suspect nucleic acid depending upon the identity of said suspect nucleic acid to form a hybridized complex; b) contacting said modified nucleic acid with a mixture of labelled antibodies, one of which is selected to complex with at least the modified portion of each of said modified nucleic acids to form a hybridized complex; and c) measuring for the presencee and/or extent of the presence of said labelled antibodies as an indication of the presence or extent of presence of at least one of said suspect nucleic acids.
Claim 57
Dependent Claim
141 words
43) A method of assaying for the presence or quantity of a suspect nucleic acid in a suspect sample characterised in that it comprises the steps of: a) contacting said suspect sample with a plurality of modified nucleic acids, each of said nucleic acids being selected such that it may hybridize with said suspect nucleic acid depending upon the identity of said suspect nucleic acid to form a hybridized complex; b) contacting said modified nucleic acid with a mixture of labelled antibodies, one of which is selected to complex with at least the modified portion of each of said modified nucleic acids to form a hybridized complex; and c) measuring for the presencee and/or extent of the presence of said labelled antibodies as an indication of the presence or extent of presence of at least one of said suspect nucleic acids.
Claim 58
Dependent Claim
23 words
44) The method as defined in claim 1 further comprising first contacting said modified nucleic acid with non-labelled antibodies prior to step b).
References Cited (Backward Citations)
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