US5281519A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Simple, rapid and reliable method for detecting thalassemia
Assignee: United States Department of Health and Human Services
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Filing: 1992-10-23
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Granted: 1994-01-25
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Expires: 2012-10-23
Abstract
Abstract A simple, rapid and reliable method for diagnosis of thalassemia is described. The method comprises amplification of the cDNA by polymerase chain reaction and determining the ratio between (alpha) and (beta) hemoglobin chain mRNAs.
Inventors
- 👤 Alan N. Schechter
- 👤 Shu-Zhen Huang
- 👤 Griffin P. Rodgers
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-03 17:59
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6876; C12Q1/6883; C12Q2600/00; C12Q2600/156
Claims Analysis
Total Claims: 9
Claim 1
Independent Claim
164 words
A method for diagnosing thalassemia, comprising the steps of: (a) extracting with an RNase inhibitor reagent an mRNA sample from the peripheral blood of an individual suspected of thalassemia; (b) preparing a cDNA sample complementary to the mRNA obtained in step (a); (c) amplifying the cDNA obtained from step (b) by polymerase chain reaction using thalassemia cDNA-specific primers selected from the group consisting of: ______________________________________
α 5'
GACAAGACCAACGTCAAGGCCGCC
α 3'
CGGAGGGACCTGTTCAAGGAC
______________________________________ for the α hemoglobin chain, and ______________________________________
β1 5' ACACAACTGTGTTCACTAGC
β1 3' AAGTGGGGTGGTCACGTCCGA
and
β4 5' ACACAACTGTGTTCACTAGC
β4 3' GCGAAAGAACGACAGGTTAAA
______________________________________ for the β hemoglobin chain and then (d) determining radiometric or densitometric ratio between α and β hemoglobin chain mRNas by the ratio between corresponding amplified cDNA obtained in step (c), a ratio greater than 100 being indicative of β o thalassemia major, a ratio in the range of 3.6 to 4.5 being indicative of β + thalassemia major and a ratio of less than 1.0 being indicative of α thalassemia.
Claim 2
Independent Claim
164 words
A method for diagnosing thalassemia, comprising the steps of: (a) extracting with an RNase inhibitor reagent an mRNA sample from the peripheral blood of an individual suspected of thalassemia; (b) preparing a cDNA sample complementary to the mRNA obtained in step (a); (c) amplifying the cDNA obtained from step (b) by polymerase chain reaction using thalassemia cDNA-specific primers selected from the group consisting of: ______________________________________
α 5'
GACAAGACCAACGTCAAGGCCGCC
α 3'
CGGAGGGACCTGTTCAAGGAC
______________________________________ for the α hemoglobin chain, and ______________________________________
β1 5' ACACAACTGTGTTCACTAGC
β1 3' AAGTGGGGTGGTCACGTCCGA
and
β4 5' ACACAACTGTGTTCACTAGC
β4 3' GCGAAAGAACGACAGGTTAAA
______________________________________ for the β hemoglobin chain and then (d) determining radiometric or densitometric ratio between α and β hemoglobin chain mRNas by the ratio between corresponding amplified cDNA obtained in step (c), a ratio greater than 100 being indicative of β o thalassemia major, a ratio in the range of 3.6 to 4.5 being indicative of β + thalassemia major and a ratio of less than 1.0 being indicative of α thalassemia.
Claim 3
Dependent Claim
25 words
The method of claim 1 wherein a radioactive purine or pyrimidine triphosphate base is added to facilitate quantitation of the product obtained from step (c).
Claim 4
Dependent Claim
11 words
The method of claim 2 wherein said base is B2P -dCTP.
Claim 5
Dependent Claim
26 words
The method of claim 3 wherein a radiometric ratio between α and β mRNA in the range of 14-100 is indicative of β o thalassemia major.
Claim 6
Dependent Claim
37 words
The method of claim 3 wherein a radiometric ratio between α and β mRNA in the range of 5.2-7.2 is indicative of β + thalassemia major and a range less than 1.0 is indicative of α thalassemia.
Claim 7
Independent Claim
47 words
A method for determining the therapeutic efficacy of a treatment for thalassemia, sickle cell disease and other hemoglobinopathies, comprising periodically monitoring the increases in the ratio of γmRNA to βmRNA as an index of therapeutic response, an increase in said ratio being indicative of an efficacious therapy.
Claim 8
Independent Claim
47 words
A method for determining the therapeutic efficacy of a treatment for thalassemia, sickle cell disease and other hemoglobinopathies, comprising periodically monitoring the increases in the ratio of γmRNA to βmRNA as an index of therapeutic response, an increase in said ratio being indicative of an efficacious therapy.
Claim 9
Dependent Claim
13 words
The method of claim 1 wherein the RNase inhibitor is Vanadyl Ribonucleoside Complex.
References Cited (Backward Citations)
US4683195A
Cited
US4683195B1
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1997-01-24 | 1997-07-26 | 1998-01-25 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2001-01-24 | 2001-07-26 | 2002-01-25 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2005-01-24 | 2005-07-26 | 2006-01-25 | $7700.00 | Window Closed / Lapsed |