Blennerhassett G T, Furth M E, Anderson A, Burns J P, Chaganti R S, Blick M, Talpaz M, Dev V G, Chan L C, Wiedemann L M
Oncogene Science, Inc., Manhasset, NY 11030.
Leukemia. 1988 Oct;2(10):648-57.
We report the clinical evaluation of an improved DNA probe assay for the characteristic genetic marker of human CML, observed by cytogenetics and designated the Philadelphia chromosome (Ph1). The Ph1 chromosome results from the fusion of c-abl proto-oncogene sequences from chromosome 9 to phl gene sequence on chromosome 22. (The phl gene is often referred to as bcr. However, for clarity we prefer to reserve the designation "bcr" for the region within the phl gene in which translocation breakpoints have been found to occur. We also find it useful to distinguish between two such regions in phl, bcr-210 and bcr-190, named after the 210- and 190-kDa phl/abl fusion proteins resulting from translocations with breakpoints in the respective regions. We refer to the corresponding chromosomal translocations as Ph1(bcr-210) and Ph1(bcr-190).) DNA, extracted from peripheral blood (PB) or bone marrow (BM) and digested with restriction endonuclease BglII, is hybridized with a probe (phl/bcr-3) spanning a breakpoint cluster region within phl. Rearrangements are revealed by the presence of one or two novel junction fragments. Clinical specimens from leukemic patients with active disease were compared by cytogenetic and DNA probe analysis at seven centers in the United States and Europe. The probe assay identified the phl rearrangement in 190 of 191 cases of Ph1-positive CML, as well as in 12 of 27 clinically diagnosed CML specimens lacking a typical Ph1 chromosome. DNA rearrangements also were seen in two of six cases of Ph1-positive ALL. No false positive results were obtained among 93 non-leukemic controls. Mixing experiments showed that the DNA probe assay can detect as few as 1% leukemic cells in a specimen. A preliminary study of CML patients in remission after allogeneic BM transplantation revealed a small fraction of residual Ph1-positive leukemic cells in a significant number of such patients.
我们报告了一种改进的DNA探针检测法的临床评估，该检测法用于检测人类慢性粒细胞白血病（CML）的特征性遗传标记，此标记通过细胞遗传学观察到，并被命名为费城染色体（Ph1）。Ph1染色体是由9号染色体上的c-abl原癌基因序列与22号染色体上的phl基因序列融合而成。（phl基因常被称为bcr。然而，为清晰起见，我们更倾向于将“bcr”这一名称保留给phl基因中已发现易位断点的区域。我们还发现区分phl基因中的两个此类区域很有用，即bcr-210和bcr-190，它们以分别在各自区域发生易位后产生的210-kDa和190-kDa的phl/abl融合蛋白命名。我们将相应的染色体易位称为Ph1(bcr-210)和Ph1(bcr-190)。）从外周血（PB）或骨髓（BM）中提取DNA，并用限制性内切酶BglII消化，然后与跨越phl基因内一个断点簇区域的探针（phl/bcr-3）杂交。通过一个或两个新的连接片段的存在来揭示重排情况。来自美国和欧洲七个中心的活动性疾病白血病患者的临床标本通过细胞遗传学和DNA探针分析进行了比较。探针检测法在191例Ph1阳性CML病例中的190例以及27例临床诊断为CML但缺乏典型Ph1染色体的标本中的12例中鉴定出phl重排。在6例Ph1阳性急性淋巴细胞白血病（ALL）病例中的2例中也发现了DNA重排。在93例非白血病对照中未获得假阳性结果。混合实验表明，DNA探针检测法能够检测出标本中低至1%的白血病细胞。对异基因骨髓移植后缓解期的CML患者进行的一项初步研究显示，相当数量的此类患者中存在一小部分残留的Ph1阳性白血病细胞。
