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1996 Fiscal Year Final Research Report Summary

Detection of minimal residual disease of leukemia using digoxigenin labeled probe

Research Project

Project/Area Number 07672495
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Laboratory medicine
Research InstitutionKyoto Prefectural University of Medicine

Principal Investigator

YUKOTO Shouhei  Kyoto Prefecteral University of Medicine, Faculty of Medicine, assistant professor, 医学部・, 助手 (80231687)

Co-Investigator(Kenkyū-buntansha) SONODA Yoshiaki  Kyoto Prefectural University of Medicine Faculty of Medicine, associate professo, 医学部, 講師 (60206688)
Project Period (FY) 1995 – 1996
Keywordsminimal residual disease / T cell receptor gene / immunoglobulin heavy chain gene / digoxigenin / acute lymphoblastic leukemia
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.

  • Research Products

    (21 results)

All Other

All Publications (21 results)

  • [Publications] Nakao M,Yokota sら: "Alterations of CDKN2 gene structure in childhood acute lymphoblastic leukemia:mutations of CDKN2 are observed preferentially in T lineage." Leukemia. 10. 249-254 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yasuda H,Nakao M,ら: "T-cell lymphoma presenting with pericardial and pleural effusion as the initial and primary lesion:Cytogenetic and molecular evidence." Internal Medicine. 35(2). 150-154 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Nakao M,Yokota sら: "Detection and quantitation of TEL/AML1 fusion transcripts by polymeraes chain reaction in childhood acute lymphoblastic leukemia." Leukemia. 10. 1463-1470 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Nakao M,Yokota sら: "Internal tandem duplication of the flt3 gene found in acute myeloid leukemia." Leukemia. 10. 1811-1818 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 三澤信一、谷脇雅史、横田昇平: "FISHおよびPCR法による微小残存白血病の検出と臨床応用." 臨床血液. 36. 293-297 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kaneko H et al: "Neurofibromatosis 1 gene(NF1)mutation is a raregenetic event in myelo-dysplastic syndrome regardless of the disease prognosis." Internatl J Hematol. 61. 113-116 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 三澤信一ら: "血液疾患に併発した重症感染症に対するFosfomycinm Sulbactam/Cefoperazone連続併用療法." Jpn J Antibiot. 48(4). 514-521 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 堀池重夫ら: "急性白血病治療後の顆粒球減少時における感染予防の検討-NorfloxacinとPolymyxin Bの比較-" 日化療会誌. 43(2). 207-212 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Seriu T,Yokota S: "Prospective monitoring of minimal residual disease during the course of chemotherapy in patients with acute lymphoblastic leukemia,and detection of contaminating tumor cells in peripheral blood stem cells for auto-transplantation." Leukemia. 9(4). 615-623 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] I ikawa K et al: "Detection of neoplastic clone in the hypoplastic and recovery phases preceding acute lymphoblastic leukemia by in vitro amplification of rearranged T-cell receptor δ chain gene." J.ped.Hematol.Oncol.17(3). 270-275 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 横田昇平、中尾 誠: "微小残存病変の検出との意義.カレント内科学No.7 急性白血病と悪性リンパ腫." 金原出版、東京, 6 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Nakao M,Yokotta S,Iwai T,Kaneko H,Horiike S Leukemia Kashima K,Sonoda Y,Sonoda Y,Fujimoto T,S Misawa: "Internal tandem duplication of the fit3 gene found in acute myeloid leukemia." Leukemia. 10. 1811-1818 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Nakao M,Yokota S,Horiike S,Taniwaki M,KashimaK,Sonoda Y,Koizumi S,Takaue Y,Matsushita T,Fujimoto T,Misawa S: "Detection and quantitation of TEL/AML1 fusion transcripts by polymerase chain reaction in chidhood acute lymphoblastic leukemia." Leukemia. 10. 1463-1470 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yokota S,Nakano M : (ed. by H Saitoh): "Detection of minimal residual disease and its clinical significance" Leukemia and lymphoma in Current Medicine. 7. 130-145 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yasuda H,Nakao M,kanemasa H,Ueda T,Mori T,Fujino H,Ohishi T,Inada Y, Tanigawa H,Horiike S,Yokota S,Misawa S,Kashima K.: "T-cell lymphoma presenting with pericardial and pleural effusion as the initial and primary lesion : Cytogenetic and molecular evidence." Internal Medicine. 35 (2). 150-154 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Nakao M,Yokota S,Seriu T,Koizumi S,Takeue Y,Leukemia Misawa S.: "Alterations of CDKN2 gene structure inchildhood acute lymphoblastic leukemia : mutations of CDKN2 are observed preferentially in T lineage." Leukemia. 10. 249-254 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ishikawa K,Seriu T,Watanabe A,Hayasawa K,Takeda O,Sato T,Takahashi I, Suzuki T,Nishinomiya F,Sato W,Takeda G,Shimada K,Yokota S.: "Detection of neoplastic clone in the hypoplastic and recovery phases preceding acute lymphoblastic leukemia by in vitro amplification of rearranged T-cell receptor delta chain gene." J.Ped.Hamatol.Oncol.17(3). 270-275 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yokota S,Nakao M,Seriu T: "The detection of minimal residula disease and the clinical relevance. (ed.by F Takaku, M Ogawa)" in peripheral blood stem cell transplantation. 56-60 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kaneko H,Horiike S,Nakai H,Ueda Y,Nakao M,Hirakawa K,Yokota S,Taniwaki M,Misawa S,Kashima K: "Neurofibromatosis 1 gene (NF1) mutation is a rare genetic event in myelo-dysplastic syndrome regardless of the disease prognosis." Internatl J Hematol. 61. 113-116 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Misawa S,Taniwaki M,Yokota S: "Detection of minimal residal disease by means of FISH and PCR methods and its clinical evaluation." Jpn J Clin Hematol. 36. 293-297 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Seriu T,Yokota S,Nakao M,Takue Y,Koizumi S,Kawai S,Fujimoto T: "Prospective monitoring of minimal residual disease during the course of chemotherapy in patients with acute lymphoblastic leukemia, and detection of contaminating tumor cells in peripheral blood stem cells for autotransplantation." Leukemia. 9(4). 615-623 (1995)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 1999-03-09  

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