US4731325A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Arrays of alternating nucleic acid fragments for hybridization arrays
Assignee: SANGTEC MOLECULAR DIAGNOSTICS AB
•
Filing: 1985-01-24
•
Granted: 1988-03-15
•
Expires: 2005-01-24
Abstract
Abstract The invention is related to improved nucleic acid reagents comprising arrays of nucleic acid fragments and combinations of such fragments. The preparation of such fragments by recombinant DNA techniques and their use in sandwich hybridization methods is also described. By making different combinations of the nucleic acid fragments-some labeled and some affixed to solid carriers, it is possible to create kits for the identification of e.g. veneral diseases. The improved nucleic acid reagents comprise two series of at least two alternating nucleic acid fragments, which are homologous to sequences in the nucleic acid to be identified, one of the series being labeled and one affixed to a solid carrier. Nucleic acid fragments belonging to different series must not be homologous to each other. Sandwich hybridization tests performed with arrays of nucleic acid fragments are at least four times as sensitive as sandwich hybridization tests performed with reagents belonging to the prior art.
Inventors
- 👤 Airi M. Palva
- 👤 Tuula M. Ranki
- 👤 Hans E. Soderlund
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-25 19:37
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/70; C12Q1/701; C12Q1/705; C12Q1/68; C12Q1/6813; C12Q1/6816; C12Q1/682; C12Q1/6834; C12Q1/6837; C12Q1/6876; C12Q1/6888; C12Q1/689
Claims Analysis
Total Claims: 26
Claim 1
Independent Claim
87 words
A nucleic acid reagent for use in a sandwich hybridization array to detect the identity of a nucleic acid, said nucleic acid reagent comprising at least two sets of at least two alternating nucleic acid fragments which are homologous to the nucleic acid which is to be identified, said alternating nucleic acid fragments being not homologous to one another, wherein at least one of the sets of nucleic acid fragments are labeled and at least one set of nucleic acid fragments are affixed to a solid carrier.
Claim 2
Dependent Claim
87 words
A nucleic acid reagent for use in a sandwich hybridization array to detect the identity of a nucleic acid, said nucleic acid reagent comprising at least two sets of at least two alternating nucleic acid fragments which are homologous to the nucleic acid which is to be identified, said alternating nucleic acid fragments being not homologous to one another, wherein at least one of the sets of nucleic acid fragments are labeled and at least one set of nucleic acid fragments are affixed to a solid carrier.
Claim 3
Dependent Claim
38 words
The nucleic acid reagent according to claim 21, wherein some or all of the nucleic acid fragments on one or more of the sets of nucleic acid fragments are joined to form an array of nucleic acid fragments.
Claim 4
Dependent Claim
16 words
The nucleic acid reagent according to claim 2, comprising nucleic acid fragments which have vector-derived parts.
Claim 5
Dependent Claim
50 words
The nucleic acid reagent according to claim 2, comprising the recombinant plasmid pKTH1271 or derivatives thereof, which recombinant plasmid contains the DNA of Cytomegalovirus Ad169 and is cloned into the host Escherichia coli K12 HB101, and the deposit number of said host containing the recombinant plasmid pKTH1271 is DSM 2826.
Claim 6
Dependent Claim
16 words
The nucleic acid reagent according to claim 2, comprising nucleic acid fragments which have vector-derived parts.
Claim 7
Dependent Claim
49 words
The nucleic acid reagent according to claim 1, comprising the recombinant plasmid pKTH1271 or derivatives thereof, which recombinant plasmid contains the DNA of Cytomegalovirus AD169 and is cloned into the host Escherichia coli K12 HB101, and the deposit number of said host containing the recombinant plasmid pKTH1271 DSM 2826.
Claim 8
Dependent Claim
55 words
Nucleic acid reagents comprising arrays of alternating nucleic acid fragments comprising the recombinant plasmid pKTH1220 or its derivatives and which recombinant plasmid contains the DNA of Chlamydia trachomatis L2 bacterium and is cloned into the host Escherichia coli K12, HB101, and the deposit number of this host comprising the recombinant plasmid pKTH1220 is DSM 2825.
Claim 9
Dependent Claim
55 words
Nucleic acid reagents comprising arrays of alternating nucleic acid fragments comprising the recombinant plasmid pKTH1220 or its derivatives and which recombinant plasmid contains the DNA of Chlamydia trachomatis L2 bacterium and is cloned into the host Escherichia coli K12, HB101, and the deposit number of this host comprising the recombinant plasmid pKTH1220 is DSM 2825.
Claim 10
Dependent Claim
80 words
Nucleic acid reagent comprising two or more series of at least two alternating arrays of nucleic acid fragments sufficiently homologous to the nucleic acid which is to be identified but not homologous to one another, comprising the recombinant plasmid pKTH1220 or its derivatives and which recombinant plasmid contains the DNA of Chlamydia trachomatis L2 bacterium and is cloned into the host Escherichia coli K12, HB101, and the deposit number of this host comprising the recombinant plasmid pKTH1220 is DSM 2825.
Claim 11
Dependent Claim
80 words
Nucleic acid reagent comprising two or more series of at least two alternating arrays of nucleic acid fragments sufficiently homologous to the nucleic acid which is to be identified but not homologous to one another, comprising the recombinant plasmid pKTH1220 or its derivatives and which recombinant plasmid contains the DNA of Chlamydia trachomatis L2 bacterium and is cloned into the host Escherichia coli K12, HB101, and the deposit number of this host comprising the recombinant plasmid pKTH1220 is DSM 2825.
Claim 12
Dependent Claim
45 words
A method for identifying more than one nucleic acid comprising exposing the nucleic acids to the nucleic acid reagent according to claim 2, wherein combinations of nucleic acid reagents are assembled from nucleic acid fragments which are homologous to the nucleic acids to be identified.
Claim 13
Dependent Claim
28 words
A method for the preparation of the nucleic acid reagent according to claim 2, wherein the nucleic acid reagent is prepared synthetically, semi-synthetically or by using recombinant-DNA techniques.
Claim 14
Dependent Claim
93 words
The method according to claim 10, wherein the preparation of the nuclei acid reagent comprises: (a) isolating nucleic acids which are homologous to the nucleic acids to be identified; (b) cloning said nucleic acids into vectors; (c) fragmenting said nucleic acids using restriction enzymes into at least two sets of at least two alternating nucleic acid fragments; (d) cloning said nucleic acid fragments into vectors; (e) labeling one set of nucleic acid fragments; and (f) fixing said nucleic acid fragments belonging to a second set of nucleic acid fragments to a solid carrier.
Claim 15
Dependent Claim
30 words
The method according to claim 11, wherein after step (c), some or all of the nucleic acid fragments are combined to form arrays of nucleic acids by using suitable ligases.
Claim 16
Dependent Claim
21 words
The method according to claim 11, wherein in step (d) the fragments belonging to different sets are cloned into different vectors.
Claim 17
Dependent Claim
45 words
A method for identifying more than one nucleic acid comprising exposing the nucleic acids to the nucleic acid reagent according to claim 1, wherein combinations of nucleic acid reagents are assembled from nucleic acid fragments which are homologous to the nucleic acids to be identified.
Claim 18
Dependent Claim
27 words
A method for the preparation of the nucleic acid reagent according to claim 1, wherein the nucleic acid reagent is prepared synthetically,semi-synthetically or by using recombinant-DNA techniques.
Claim 19
Dependent Claim
93 words
The method according to claim 15 wherein the preparation of the nucleic acid reagent comprises: (a) isolating nucleic acids which are homologous to the nucleic acids to be identified; (b) cloning said nucleic acids into vectors; (c) fragmenting said nucleic acids using restriction enzymes into at least two sets of at least two alternating nucleic acid fragments; (d) cloning said nucleic acid fragments into vectors; (e) labeling one set of nucleic acid fragments; and (f) fixing said nucleic acid fragments belonging to a second set of nucleic acid fragments to a solid carrier.
Claim 20
Dependent Claim
21 words
The method according to claim 16, wherein in step (d) the fragments belonging to different sets are cloned into different vectors.
Claim 21
Independent Claim
115 words
A method for conducting a sandwich hybridization array using a nucleic acid reagent to detect the identity of a nucleic acid, said method comprising exposing said nucleic acid to said nucleic acid reagent, said nucleic acid reagent comprising at least two sets of at least two alternating nucleic acid fragments which are homologous to the nucleic acid which is to be identified, said nucleic acid fragments being not homologous to one another, wherein the nucleic acid fragments from each of said sets alternate in said nucleic acid reagent and at least one of said sets of nucleic acid fragments are labeled and at least one set of nucleic fragments are affixed to a solid carrier.
Claim 22
Independent Claim
115 words
A method for conducting a sandwich hybridization array using a nucleic acid reagent to detect the identity of a nucleic acid, said method comprising exposing said nucleic acid to said nucleic acid reagent, said nucleic acid reagent comprising at least two sets of at least two alternating nucleic acid fragments which are homologous to the nucleic acid which is to be identified, said nucleic acid fragments being not homologous to one another, wherein the nucleic acid fragments from each of said sets alternate in said nucleic acid reagent and at least one of said sets of nucleic acid fragments are labeled and at least one set of nucleic fragments are affixed to a solid carrier.
Claim 23
Dependent Claim
36 words
The method according to claim 18, wherein some or all of the nucleic acid fragments on one or more of the sets of nucleic acid fragments are joined ot form an array of nucleic acid fragments.
Claim 24
Dependent Claim
14 words
The method according to claim 18, comprising nucleic acid fragments which have vector-derived parts.
Claim 25
Dependent Claim
50 words
The method according to claim 18, comprising the recombinant plasmid pKTH1220 or derivatives thereof, which recombinant plasmid contains the DNA of Chlamydia trachomatis L2 bacterium and is cloned into the host Escherichia coli K12 HB101, and the deposit number of said host containing the recombinant plasmid pKTH1220 is DSM 2825.
Claim 26
Dependent Claim
49 words
The method according to claim 18, comprising the recombinant plasmid pKTH1271 or derivatives thereof, which recombinant plasmid contains the DNA of Cytomegalovirus AD169 and is cloned into the host Escherichia coli K 12 HB101, and the eposit number of said host containing the recombinant plasmid KTH1271 is DSM 2826.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1991-03-15 | 1991-09-14 | 1992-03-15 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1995-03-15 | 1995-09-14 | 1996-03-15 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1999-03-15 | 1999-09-14 | 2000-03-15 | $7700.00 | Window Closed / Lapsed |