US4302437A
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Mitogen stimulated lymphocyte transformation
Assignee: Individual
•
Filing: 1979-08-30
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Granted: 1981-11-24
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Expires: 1999-08-30
Abstract
Abstract Mitogen stimulated transformation of blood lymphocytes is effected with a whole blood sample of a quantity of no greater than 0.5 ml, and a mitogen to whole blood sample ratio which prevents agglutination of blood cells to thereby facilitate uniform uptake of reagents by the lymphocytes in subsequent testing. Lymphocyte transformation can be measured by use of a radioisotope, and the technique is particularly suitable for measuring vitamin deficiency by a dU or thymidine suppression test, or for determining lymphocytic malignancy. The use of a whole blood culture eliminates lymphocyte separation, and the use of micro quantities of blood extends clinical application to routine laboratories and pediatric patients.
Inventors
- 👤 Victor Herbert
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 14:17
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/82; G01N33/58; G01N33/60; Y; Y10; Y10T; Y10T436/00; Y10T436/10; Y10T436/101666
Claims Analysis
Total Claims: 33
Claim 1
Independent Claim
84 words
In a process for determining vitamin deficiency by a deoxyuridine suppression test wherein a blood sample is preincubated with a mitogen to stimulate transformation of blood lymphocytes to produce DNA, followed by incubation with deoxyuridine and tracer, the improvement comprising: employing a whole blood sample of a quantity of no greater than 0.5 ml, and mitogen in an amount to stimulate transformation of lymphocytes in the whole blood sample and provide a mitogen to whole blood sample ratio which prevents agglutination of blood cells.
Claim 2
Dependent Claim
84 words
In a process for determining vitamin deficiency by a deoxyuridine suppression test wherein a blood sample is preincubated with a mitogen to stimulate transformation of blood lymphocytes to produce DNA, followed by incubation with deoxyuridine and tracer, the improvement comprising: employing a whole blood sample of a quantity of no greater than 0.5 ml, and mitogen in an amount to stimulate transformation of lymphocytes in the whole blood sample and provide a mitogen to whole blood sample ratio which prevents agglutination of blood cells.
Claim 3
Dependent Claim
24 words
The process of claim 1 wherein the incubation with deoxyuridine is conducted after mitogen stimulation for a period sufficient to induce peak mitogen response.
Claim 4
Dependent Claim
21 words
The process of claim 1 wherein the incubation with deoxyuridine is conducted after mitogen stimulation for a period of four days.
Claim 5
Dependent Claim
13 words
The process of claim 1 wherein the tracer is ( 125 I) deoxyuridine.
Claim 6
Dependent Claim
18 words
The process of claim 4 wherein the tracer and sample are incubated for from 1 to 4 hours.
Claim 7
Dependent Claim
13 words
The process of claim 1 wherein the tracer is ( 3 H) thymidine.
Claim 8
Dependent Claim
18 words
The process of claim 6 wherein the tracer and sample are incubated for from 1 to 4 hours.
Claim 9
Dependent Claim
14 words
The process of claim 1 wherein the whole blood sample quantity is 0.1 ml.
Claim 10
Dependent Claim
23 words
The process of claim 8 wherein the mitogen is employed in an amount of 20 μg per 0.1 ml of whole blood sample.
Claim 11
Dependent Claim
10 words
The process of claim 9 wherein the mitogen is PHA.
Claim 12
Dependent Claim
13 words
The process of claim 10 wherein the tracer is ( 125 I) deoxyuridine.
Claim 13
Dependent Claim
13 words
The process of claim 10 wherein the tracer is ( 3 H) thymidine.
Claim 14
Dependent Claim
82 words
In a process for determining vitamin deficiency by a thymidine suppression test wherein a blood sample is preincubated with a mitogen to stimulate transformation of blood lymphocyte to produce DNA, followed by incubation with thymidine and tracer, the improvement comprising: employing a whole blood sample of a quantity of no greater than 0.5 ml and mitogen in an amount to stimulate transformation in the whole blood sample and provide a mitogen to whole blood sample ratio which prevents agglutination of blood cells.
Claim 15
Dependent Claim
82 words
In a process for determining vitamin deficiency by a thymidine suppression test wherein a blood sample is preincubated with a mitogen to stimulate transformation of blood lymphocyte to produce DNA, followed by incubation with thymidine and tracer, the improvement comprising: employing a whole blood sample of a quantity of no greater than 0.5 ml and mitogen in an amount to stimulate transformation in the whole blood sample and provide a mitogen to whole blood sample ratio which prevents agglutination of blood cells.
Claim 16
Dependent Claim
23 words
The process of claim 13 wherein incubation with thymidine is conducted after mitogen stimulation for a period sufficient to induce peak mitogen response.
Claim 17
Dependent Claim
20 words
The process of claim 13 wherein incubation with thymidine is conducted after mitogen stimulation for a period of four days.
Claim 18
Dependent Claim
13 words
The process of claim 13 wherein the tracer is ( 125 I) deoxyuridine.
Claim 19
Dependent Claim
14 words
The process of claim 13 wherein the whole blood sample quantity is 0.1 ml.
Claim 20
Dependent Claim
22 words
The process of claim 17 wherein the mitogen is employed in an amount of 20 μg per 0.1 ml of whole blood.
Claim 21
Dependent Claim
10 words
The process of claim 18 wherein the mitogen is PHA.
Claim 22
Dependent Claim
83 words
In a process for determining lymphocytic malignancy by determining delayed mitogen stimulated lymphocyte transformation, wherein a blood sample is preincubated with a mitogen to stimulate transformation of blood lymphocytes to produce DNA followed by incubation with a tracer, the improvement comprising: employing a whole blood sample of a quantity of no greater than 0.5 ml and mitogen in an amount to stimulate transformation in the whole blood sample and provide a mitogen to whole blood sample ratio which prevents agglutination of blood cells.
Claim 23
Dependent Claim
83 words
In a process for determining lymphocytic malignancy by determining delayed mitogen stimulated lymphocyte transformation, wherein a blood sample is preincubated with a mitogen to stimulate transformation of blood lymphocytes to produce DNA followed by incubation with a tracer, the improvement comprising: employing a whole blood sample of a quantity of no greater than 0.5 ml and mitogen in an amount to stimulate transformation in the whole blood sample and provide a mitogen to whole blood sample ratio which prevents agglutination of blood cells.
Claim 24
Dependent Claim
14 words
The process of claim 20 wherein the whole blood sample quantity is 0.1 ml.
Claim 25
Dependent Claim
22 words
The process of claim 21 wherein the mitogen is employed in an amount of 20 μg per 0.1 ml of whole blood.
Claim 26
Dependent Claim
10 words
The process of claim 22 wherein the mitogen is PHA.
Claim 27
Dependent Claim
32 words
The process of claim 1 wherein the deoxyuridine suppression test is effected by incubation with deoxyuridine in an amount of less than 5 μmol, based on 1 ml of total blood culture.
Claim 28
Dependent Claim
29 words
The process of claim 24 wherein the amount of deoxyuridine is at least 1 μmol and no greater than 3 μmol, based on 1 ml of total blood culture.
Claim 29
Dependent Claim
30 words
The process of claim 11 wherein the deoxyuridine suppression test is effected by incubation with deoxyuridine in an amount of 2 μmol, based on 1 ml of total blood culture.
Claim 30
Dependent Claim
29 words
The process of claim 13 wherein the amount of thymidine is at least 1 μmol and no greater than 3 mol, based on 1 ml of total blood culture.
Claim 31
Dependent Claim
26 words
The process of claim 1 wherein the mitogen to whole blood sample ratio is no greater than 40 μg per 0.1 ml of whole blood sample.
Claim 32
Dependent Claim
26 words
The process of claim 13 wherein the mitogen to whole blood sample ratio is no greater than 40 μg per 0.1 ml of whole blood sample.
Claim 33
Dependent Claim
26 words
The process of claim 20 wherein the mitogen to whole blood sample ratio is no greater than 40 μg per 0.1 ml. of whole blood sample.
References Cited (Backward Citations)
No citation records indexed.
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1984-11-23 | 1985-05-25 | 1985-11-24 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1988-11-23 | 1989-05-25 | 1989-11-24 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1992-11-23 | 1993-05-25 | 1993-11-24 | $7700.00 | Window Closed / Lapsed |