悪性リンパ腫の微量病変の検出

恶性淋巴瘤微小病变的检测

• 批准号: 07772250
• 负责人: 星野 敦
• 金额: $ 0.51万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Encouragement of Young Scientists (A)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 无数据
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07772250/
• 关键词: 悪性リンパ腫; 微量残存病変; DNA診断; PCR; 免疫グロブリン重鎖遺伝子再構成; 骨髄浸潤; 臨床病期

悪性リンパ腫における,骨髄・末梢血への微量病変の浸潤を,分子生物学的手法を用いて高感度に(検出する目的で研究を行った.17例のB細胞系非ホジキンリンパ腫について行った.リンパ節生検材料から,genomic DNAを抽出し,免疫グロブリン重鎖遺伝子(immunoglobulin heavy chain:IgH)のgene rearrangementの検出をPCRにて行った.17例中,11例でrearrangement bandが検出された.得られたrearrangement bandをポリアクリルアミドゲルにて分画,精製し、T-vectorにライゲーションし,シークエンスを行った.これをもとに,各症例毎に,リンパ腫細胞DNAに特異的なPCRプライマーと非RIプローブを作成した.微量病変の検出を目的とした検体(骨髄穿刺液,末梢血など)からDNAを抽出した.各症例に特異的なプライマーを用いてPCRを行い,腫瘍細胞特異的DNAの増幅を行った.さらに非RIプローブを用てサザン・プロット解析を行い,検体中の腫瘍細胞特異的DNAの検出を行った.11例中9例で微量病変が骨髄,末梢血中で検出された.これらは全例で,従来の光顕レベルではリンパ腫細胞の検出が不可能であった.これにより臨床的病期診断がIV期に変更されると考えられた症例は6例であった.(他3例は画像診断などで臨床病期がはじめからIV期と診断されていた.)このことから悪性リンパ腫における微量病変のDNA診断が,臨床的に有用であると思われる.今後症例を増やして,予後との関連などをさらに追求して行く予定である.また,得られたDNAプローブを用いたin situ hybridezation(ISH)によるsingle cellレベルでの検出は,現在,検出条件を検討中である.

The method of molecular biology is to use the technique of molecular biology to study the disease with high sensitivity. 17 cases of B cell line were not infected. The raw material was extracted from the raw material, the genomic DNA was extracted, and the immunized mice were immunized with immunoglobulin heavy chain:IgH, gene rearrangement, PCR. Of the 17 cases, 11 were diagnosed with rearrangement band. Please tell me that the pictures are divided, that is, the rearrangement band, the T-vector, and the lines. This is not the case of the disease. This is the case of each case. This is the case of the special DNA cell. This is not the case of the PCR patient. The micro-disease was used to extract the body (bone puncture fluid, peripheral blood) and DNA. In each case, the specific PCR line was used, and the DNA width of the cell was used. In the case of non-RI diseases, the DNA of specific cells in the body was detected. Of the 11 cases, there were 9 cases of micronutrient disease, which were detected in the peripheral blood. In the case of the whole case, please tell me that it is impossible to tell you that it is impossible. The disease stage of the bed was cut off, and the IV period was changed. There were 6 cases of illness. (his portrait was cut off in 3 cases, and the bed illness stage was broken. The IV period was broken.) I don't know if I have sex, I don't know, I don't know. In the future, there will be a case of illness, and in the future, you will be in the pursuit of a predetermined situation. Now, you have to use the in situ hybridezation (ISH), the single cell, the single cell, the output. Now, you can find out the conditions.