论文部分内容阅读
对急性白血病AP-PCRDNA指纹图改变及其在急性白血病微小残留病诊断中的应用为本文研究目的。方法:以31例急性白血病患儿为研究对象,取其骨髓或外周血标本,先用Percoll不连续密度梯度离心法分离出白血病细胞,以自身正常中性粒细胞为对照,分别选用3种引物AP1、AP2、AP3进行AP-PCR扩增,每例标本均重复一次,产物经变性聚丙烯酸胺凝胶电泳后银染显色,分析结果。结果:(1)在一定的反应条件下,所有标本均扩增出清晰条带,并且有很好的重复性;(2)58.1%(18/31)可见白血病细胞相对于成熟粒细胞的指纹图改变。图谱改变包括新增带、带缺失\带密度改变。(3)AP1、AP2、AP3对所有31例标本异常检出率分另。1为41.9%、35.5%、38.7%,三种引物共检出18例异常,总检出率为58.1%。结论:AP-PCR是一种重复性好、简便易行的DNA指纹图技术,具有检出白血病细胞基因组微小改变的能力。有望成为检测微小残留病的一种新方法。
AP-PCR DNA fingerprinting of acute leukemia and its application in the diagnosis of acute leukemia minimal residual disease for the purpose of this study. Methods: Totally 31 children with acute leukemia were selected as the research objects. The bone marrow or peripheral blood samples were collected. Leukemic cells were separated by Percoll discontinuous density gradient centrifugation. Three kinds of primers AP1, AP2, AP3 for AP-PCR amplification, each specimen were repeated, the product was denatured polyacrylamide gel electrophoresis silver staining, the analysis results. Results: (1) Under certain reaction conditions, clear bands were amplified in all the specimens with good repeatability; (2) 58.1% (18/31) showed that the percentage of leukemic cells relative to mature granulocytes Fingerprint change. Pattern changes include new bands, with missing with density changes. (3) AP1, AP2, AP3 all 31 cases of abnormal detection rate of the other points. 1 were 41.9%, 35.5% and 38.7% respectively. A total of 18 abnormalities were detected by the three primers, with a total detection rate of 58.1%. Conclusion: AP-PCR is a repetitive, simple and easy DNA fingerprinting technique with the ability to detect small changes in the genome of leukemia cells. Is expected to become a new method of detecting minimal residual disease.