| Project/Area Number |
07672495
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Laboratory medicine
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| Research Institution | Kyoto Prefectural University of Medicine |
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Principal Investigator |
YUKOTO Shouhei Kyoto Prefecteral University of Medicine, Faculty of Medicine, assistant professor, 医学部・, 助手 (80231687)
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| Co-Investigator(Kenkyū-buntansha) |
SONODA Yoshiaki Kyoto Prefectural University of Medicine Faculty of Medicine, associate professo, 医学部, 講師 (60206688)
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| Project Period (FY) |
1995 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1996: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1995: ¥1,100,000 (Direct Cost: ¥1,100,000)
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| Keywords | minimal residual disease / T cell receptor gene / immunoglobulin heavy chain gene / digoxigenin / acute lymphoblastic leukemia / 免疫グロブリン重鎖遺伝子 |
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| Research Abstract |
Relapse is still the major cause of treatment failure of leukemia. The recurrence occurs through the proliferation of residual leukemic cells that are not completely eradicated by the therapy. Therefore the diagnosis of minimal residual disease (MRD) are valuable for the treatment of leukemias. To introduce the diagnosis of MRD into the laboratories of ordinary hospitals, we established the non-radioisotopic detection methods.
we used the junctional sequences of recombined T-cell receptor delta and gamma chain genes as the clonal markers. The amplified junctional PCR products were purified and labeled by digoxigenin. CR materials of the patients were amplified, blotted on the nylon membranes, and hybridized by the patient-specific probes. Using this method, we could detect one leukemic cells among 10^4 to 10^5 normal cells. We provided the labeled probes to the laboratories of same hospitals so that they could evaluate the MRD immediately by themselves.
In order to simplify the procedure, we then established another PCR-mediated mediated method using the junctional sequences as PCR primers. In this occasion, the junctional region was sequenced in each patient and the specific primers was synthesized. The primer was sent to the laboratory and used for the MRD detection. Using this method, we could detect one leukemic cell among 10^4 normal cells.
We analyzed MRD in ALL patients treated with the conventional chemotherapy. Our findings showed that gradual increase or consecutive positive results of MRD of BM was implicated in impending marrow relapse. We also qualified the hemopoietic stem-cell grafts from ALL patients. MRD was more frequently detected in the stem-cell grafts of the patients who eventually relapsed than those maintaining CR.
The diagnosis of MRD based on PCR is applicable to most of ALL children. Detection of MRD will offer a prospective view to improve the prognosis of the leukemic patients treated by modern chemotherapy.
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