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Polymerase chain reaction based assay to detect allelic loss in human DNA: loss of β-interleron gene in chronic myelogeneous leukemia

机译:基于聚合酶链反应的检测人类DNA等位基因缺失的方法:慢性粒细胞白血病中β-interleron基因的缺失

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We have developed a polymerase chain reaction (PCR) based technique to detect allelic loss. In this differential PCR a target gene and a reference gene are coamplified In the same reaction vessel. The ratio of the intensity of the two resultant bands is an Indication of relative gene dosage. This procedure is sensitive in that gene copy ratios of 2:1 and 3:2 (reference: target gene) can readily be detected. Using this differential PCR, we have examined 64 cases of chronic myelogenous leukemia (CML) for the loss of the β1-interferon gene, a relatively common event in certain human leukemias and lymphomas (1). Only one patient who was Philadelphia chromosome positive and who was in blast crisis exhibited allelic loss of the fl-interferon gene. Thus despite deletions at the β-interferon locus in the CML cell line, K562, this perturbation is rarely seen in primary CML samples.
机译:我们已经开发了一种基于聚合酶链反应(PCR)的技术来检测等位基因缺失。在该差异PCR中,靶基因和参考基因在同一反应容器中被共扩增。两个合成条带的强度之比是相对基因剂量的指示。该步骤很敏感,因为可以很容易地检测出2:1和3:2的基因复制率(参考:目标基因)。使用这种差异PCR,我们检查了64例慢性骨髓性白血病(CML)的β1-干扰素基因的丢失,这在某些人类白血病和淋巴瘤中是相对常见的事件(1)。仅有一名费城染色体阳性且处于原始危机中的患者表现出fl-干扰素基因的等位基因缺失。因此,尽管在CML细胞系K562的β-干扰素基因座处缺失,这种干扰在原代CML样品中很少见。

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