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组织学、免疫学和细胞遗传学的多参数分析是急性白血病诊断最常用的方法,其中细胞遗传学诊断可以肯定患者的预后情况,可作为肿瘤的预测工具。作者建立了一种多重RT-PCR方法扩增染色体易位所形成的嵌合基因,能同时测出急性淋巴细胞白血病(ALL)和急性粒细胞白血病(AML)的29种易位和染色体畸变。急性白血病已发现50个以上的不同易位,其中许多易位与白血病和淋巴瘤的特殊亚型有关。该方法以靠近白血病易位断裂点的结合序列为引物。引物设计成适用于所有PCR反应条件用以扩增与白血病易位相关的mRNA。逆转录合成cDNA时选用特异引物与mRNA结合,而不用随机引物或poly-dT引物,其敏感性比随机六聚引物高25～125倍。每种易位的引物经设计并优化后,可将多重RT-PCR反应减少到只用平
Multiparametric analysis of histology, immunology and cytogenetics is the most commonly used method for the diagnosis of acute leukemia. Cytogenetic diagnosis can confirm the prognosis of patients and can be used as a tumor prediction tool. The authors established a multiplex RT-PCR method to amplify the chimeric gene formed by chromosomal translocation, and can simultaneously detect 29 translocations and chromosomal aberrations in acute lymphoblastic leukemia (ALL) and acute myeloid leukemia (AML). More than 50 different translocations have been found in acute leukemia, many of which are associated with specific subtypes of leukemia and lymphoma. This method uses primers that are close to the binding sequence of the leukemia translocation breakpoints. The primers were designed to be suitable for all PCR reaction conditions to amplify mRNAs associated with leukemia translocation. When reverse transcribed cDNA was synthesized, specific primers were used to bind to mRNA instead of random primers or poly-dT primers, which were 25 to 125 times more sensitive than random hexamer primers. Each translocated primer is designed and optimized to reduce multiplex RT-PCR reactions to only