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US4582788A
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HLA typing method and cDNA probes used therein
Assignee: Hoffmann La Roche Inc
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Filing: 1983-01-07
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Granted: 1986-04-15
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Expires: 2003-01-07
Abstract
Abstract HLA typing based on restriction length polymorphism is carried out by: digesting an individual's HLA DNA with a restriction endonuclease that produces a polymorphic digestion pattern with HLA DNA; subjecting the digest to genomic blotting using a labeled cDNA hybridization probe that is complementary to an HLA DNA sequence involved in the polymorphism; and comparing the resulting genomic blotting pattern with a standard. This technique may be adapted to make paternity or transplant or transfusion compatibility determinations or to make disease association correlations to diagnose diseases or predict susceptibility to diseases. Locus specific cDNA hybridization probes, particularly probes for genes of Class II loci (D and DR loci), for use in the typing procedure are described.
Inventors
- 👤 Henry A. Erlich
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-29 18:18
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6813; C12Q1/6827; C12Q1/683; C07; C07K; C07K14/00; C07K14/435; C07K14/705; C07K14/70503; C07K14/70539; C12Q1/6876; C12Q1/6881; C12Q2600/00; C12Q2600/156; C12Q2600/172
Claims Analysis
Total Claims: 14
Claim 1
Independent Claim
98 words
An HLA typing method based on HLA DNA restriction fragment length polymorphism comprising: (a) digesting genomic HLA DNA from an individual with a restriction endonuclease that produces a polymorphic digestion pattern of a class II HLA DNA locus; (b) subjecting the digest of (a) to genomic blotting using a labeled cDNA that is complementary to a class II HLA DNA locus sequence involved in the polymorphism; and (c) comparing the genomic blotting pattern obtained in (b) with a standard genomic blotting pattern for said HLA DNA sequence obtained using said restriction endonuclease and an equivalent labeled cDNA probe.
Claim 2
Dependent Claim
98 words
An HLA typing method based on HLA DNA restriction fragment length polymorphism comprising: (a) digesting genomic HLA DNA from an individual with a restriction endonuclease that produces a polymorphic digestion pattern of a class II HLA DNA locus; (b) subjecting the digest of (a) to genomic blotting using a labeled cDNA that is complementary to a class II HLA DNA locus sequence involved in the polymorphism; and (c) comparing the genomic blotting pattern obtained in (b) with a standard genomic blotting pattern for said HLA DNA sequence obtained using said restriction endonuclease and an equivalent labeled cDNA probe.
Claim 3
Dependent Claim
20 words
The method of claim 1 wherein the locus is the DRα locus, a DRα related locus, or the DRβ locus.
Claim 4
Dependent Claim
57 words
The method of claim 1 wherein: (i) the locus is the DRα locus and the restriction endonuclease is Bg1II or EcoRV, or (ii) the locus is a DR? related locus and the restriction endonuclease is Bg1II, or (iii) the locus is the DR? locus or a DR? related locus and the restriction endonuclease is EcoRI or Bg1II.
Claim 5
Independent Claim
102 words
A method of determining whether an individual is susceptible to a disease comprising: (a) digesting genomic HLA DNA from the individual with a restriction endonuclease that produces a polymorphic digestion pattern of a class II HLA locus that is associated with the disease; (b) subjecting the digest of (a) to genomic blotting using a labeled cDNA probe that is complementary to the DNA sequence of said class II HLA locus; (c) comparing the genomic blotting pattern obtained in (b) with a standard genomic blotting pattern of an individual having said disease obtained using said restriction endonuclease and an equivalent labeled cDNA probe.
Claim 6
Independent Claim
102 words
A method of determining whether an individual is susceptible to a disease comprising: (a) digesting genomic HLA DNA from the individual with a restriction endonuclease that produces a polymorphic digestion pattern of a class II HLA locus that is associated with the disease; (b) subjecting the digest of (a) to genomic blotting using a labeled cDNA probe that is complementary to the DNA sequence of said class II HLA locus; (c) comparing the genomic blotting pattern obtained in (b) with a standard genomic blotting pattern of an individual having said disease obtained using said restriction endonuclease and an equivalent labeled cDNA probe.
Claim 7
Dependent Claim
12 words
The method of claim 4 wherein said locus is the DRα locus.
Claim 8
Dependent Claim
11 words
The method of claim 5 wherein the restriction endonuclease is Bg1II.
Claim 9
Independent Claim
124 words
A method for determining the paternity of an individual based on HLA DNA restriction fragment length polymorphism comprising: (a) digesting genomic HLA DNA of the mother of the individual, the suspected father of the individual, and the individual with a restriction endonuclease that produces a polymorphic digestion pattern of a class II HLA DNA locus; (b) subjecting each of the digests of (a) to genomic blotting using a labeled cDNA probe that is complementary to a class II HLA DNA sequence involved in the polymorphism; and (c) comparing the genomic blotting patterns obtained in (b) to determine correspondence between the individual's pattern and the mother's pattern and suspected father's pattern and thereby determining whether the suspected faster is the actual father of the individual.
Claim 10
Independent Claim
124 words
A method for determining the paternity of an individual based on HLA DNA restriction fragment length polymorphism comprising: (a) digesting genomic HLA DNA of the mother of the individual, the suspected father of the individual, and the individual with a restriction endonuclease that produces a polymorphic digestion pattern of a class II HLA DNA locus; (b) subjecting each of the digests of (a) to genomic blotting using a labeled cDNA probe that is complementary to a class II HLA DNA sequence involved in the polymorphism; and (c) comparing the genomic blotting patterns obtained in (b) to determine correspondence between the individual's pattern and the mother's pattern and suspected father's pattern and thereby determining whether the suspected faster is the actual father of the individual.
Claim 11
Independent Claim
114 words
A method for determining transplant or transfusion compatibility based on HLA DNA restriction fragment length polymorphism comprising: (a) digesting genomic HLA DNA of the transplant or transfusion donor and the transplant or transfusion host with a restriction endonuclease that produces a polymorphic digestion pattern of a class II HLA DNA locus; (b) subjecting each of the digests of (a) to genomic blotting using a labeled cDNA probe that is complementary to a class II HLA DNA sequence involved in the polymorphism; and (c) comparing the genomic blotting patterns obtained in (b) to determine correspondence therebetween and thereby determining whether the transplant or transfusion donor's HLA is compatible with the transplant or transfusion host's HLA.
Claim 12
Independent Claim
114 words
A method for determining transplant or transfusion compatibility based on HLA DNA restriction fragment length polymorphism comprising: (a) digesting genomic HLA DNA of the transplant or transfusion donor and the transplant or transfusion host with a restriction endonuclease that produces a polymorphic digestion pattern of a class II HLA DNA locus; (b) subjecting each of the digests of (a) to genomic blotting using a labeled cDNA probe that is complementary to a class II HLA DNA sequence involved in the polymorphism; and (c) comparing the genomic blotting patterns obtained in (b) to determine correspondence therebetween and thereby determining whether the transplant or transfusion donor's HLA is compatible with the transplant or transfusion host's HLA.
Claim 13
Dependent Claim
110 words
An HLA typing method based on HLA DNA restriction fragment length polymorphism comprising: (a) digesting genomic HLA DNA from an individual with a restriction endonuclease that produces a polymorphic digestion pattern of a class II HLA DNA; (b) solution hybridizing the digest of step (a) with a labeled cDNA probe that is complementary to a class II HLA DNA sequence involved in the polymorphism; (c) resolving the hybridizate of step (b); (d) detecting labeled hybrids in the resolved hybridizate; and (e) comparing the resolution pattern of the labeled hybrids with a standard resolution pattern for said HLA DNA sequence obtained using the restriction endonuclease and an equivalent labeled cDNA probe.
Claim 14
Dependent Claim
110 words
An HLA typing method based on HLA DNA restriction fragment length polymorphism comprising: (a) digesting genomic HLA DNA from an individual with a restriction endonuclease that produces a polymorphic digestion pattern of a class II HLA DNA; (b) solution hybridizing the digest of step (a) with a labeled cDNA probe that is complementary to a class II HLA DNA sequence involved in the polymorphism; (c) resolving the hybridizate of step (b); (d) detecting labeled hybrids in the resolved hybridizate; and (e) comparing the resolution pattern of the labeled hybrids with a standard resolution pattern for said HLA DNA sequence obtained using the restriction endonuclease and an equivalent labeled cDNA probe.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1989-04-14 | 1989-10-14 | 1990-04-15 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1993-04-14 | 1993-10-14 | 1994-04-15 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1997-04-14 | 1997-10-14 | 1998-04-15 | $7700.00 | Window Closed / Lapsed |