白血病微小残留病Initial Therapy of AML Diagnosis• Complete remission (CR) must be attained to cure patient• CR defined as:—Clearance of peripheral blood & bone marrow of leukemic blasts—Reconstitution of normal hematopoiesis—Resolution of leukemic infiltratesRemissionInductionPost-remission TherapyConsolidationChemotherapySCT定义Minimal residual disease(MRD)--- 微小残留病 指白血病诱导化疗完全缓解后或者骨髓移植后残留在体内少 量白血病细胞的状态, 可能是导致白血病复发的根源白血病细胞负荷 初次诊断 --- 1012 形态学缓解 --- 1010 以下 分子学缓解 --- 约106-107Acute leukemia and CML Concept of minimal residual disease (MRD)10121010106Cell numberTimeMorphological detection limitPCR detection limitI检测MRD 的意义根据细胞负荷调整缓解后化疗方案更早发现药物耐药性更早预测白血病复发评价骨髓移植净化效果为实验研究提供依据MRD 主要检测方法一、染色体核型分析二、荧光原位杂交--- FISH三、流式细胞仪---FCM四、聚合酶链反应技术---PCR五、其他:细胞培养技术,免疫荧光等染色体核型分析常用染色体显带技术检测出染色体畸变, 但其成功率依赖于标本中分裂象细胞的数量多少,从而使得检测结果各不相同,且敏感度较低,约1 :20 FISH fluorescence in situ hybridization 用荧光素标记染色体特异性探针和特异性基因的DNA 探针,通过与间期细胞核DNA 或中期染色体DNA 原位杂交识别染色体数目和结构的异常,可以进行定性、定位和相对定量分析 。优点:FISH 具有安全、快速、灵敏度高、探针能长期保存、能同时显示多种颜色等优点, 不但能显示中期分裂相, 还能显示于间期核. Interphase and metaphase FISH demonstrating a variant of the Philadelphia chromosome. The genes that are rearranged are: BCR(green on chromosome 22)/ABL (orange on chromosome 9)The Ph’ translocation appears as dual fusion (yellow) signals. Karyotype showing the Philadelphia chromosome.46,XX,t(9;22)(q34.1;q11.2)Figure 5: Detection of PML-RARa gene fusion by interphase FISH:Left: S-FISH (one wild type PML, one wild t...