非同位素的PCR-RNA转录本分子杂交法检测急淋微小残留病  

Detection of Minimal Residual Disease in Acute Lymphoblastic Leukemia Using Non-radiological PCR-RNA Molecule Hybridization

作　　者：肖燕[1] 费洪宝[1] 梁欣荃[1] 刘树茂[1] 杨爱德[1] 

机构地区：[1]同济医科大学附属协和医院儿科,武汉430022

出　　处：《同济医科大学学报》1998年第6期467-469,480,共4页Acta Universitatis Medicinae Tongji

基　　金：国家自然科学基金资助项目(No.39470786)

摘　　要：以T细胞抗原受体γ链重排基因（TCRγ）为标志,运用分别与V区和J区片段的保守序列一致,并带有T_7RNA聚合酶启动子序列的引物,将急性淋巴细胞白血病（ALL）初诊期的骨髓标本进行PCR（聚合酶链反应）.并将其产物转录成为RNA,以该RNA为探针,与ALL缓解期骨髓标本DNA的PCR产物所转录的 RNA杂交,杂交体用RNA酶消化,消化产物行聚丙烯酰酰凝胶电泳（PAGE）,尔后硝酸银染色观察检测结果。实验表明:该方法特异性强.灵敏度可达10^(-5)水平.具有快速、经济、没有同位素污染的优点,便于临床运用。The T-cell receptor gene γ-chain rearrangements(TCRγ) served as marker genes. The prime, Which was consistent with one consensus V segment and another consensus J segment separately and the promoter T7RNA polymerase was used to perform polymerase chain reaction (PCR) on first diagnostic bone marrow specimens from acute lymphoblastic leukemia (ALL). The amplified product was transcribed into a RNA probe, which was hybridized with RNA transcribed from the hybridized products,and the digested product was then PCR product of DNA in the bone marrow specimens the remission stage of All. Rnase was used to digestdetected by means of polyacrylamide gel electrophoresis (PAGE) and underwent AgNO3 staining. The results demonstrated that this assay is highly specific with the sensitivity being 10-5 level and has the advantages of rapidity,e-conomy and non-radioactivity. It is convenient to use in clinic.

关 键 词：白血病 微小残留病 ALL 聚合酶链反应 RNA杂交 

分 类 号：R733.710.4[医药卫生—肿瘤]