Qian X H, Yang A D, Fei H B, Wang C C
Department of Dediatrics, Xiehe Hospital, Tongji Medical University, Wuhan.
J Tongji Med Univ. 1993;13(3):129-33. doi: 10.1007/BF02886502.
The bcr/abl fusion gene in 20 patients with chronic myeloid leukemia (CML) was detected by RNA polymerase chain reaction, which used mRNA as the starting material to generate cDNA with reverse transcriptase followed by PCR amplification (RNA/PCR). Amplification of a sequence spanning the bcr/abl junction region was achieved by using peripheral blood cells as the source of mRNA from all 20 patients with CML, including 3 cases of Ph (-) CML, and cell line K562 was derived from patients with CML. No amplification was seen when mononuclear cells from 3 normal individuals, 2 patients with lymphoma and cell line HL-60 were used. The presence or absence of bcr exon 3 in the fusion mRNA was determined by the size of the amplified fragments. Of the 20 CML patients, 15 showed only the 165-bp amplified band (indicating retention of bcr exon 3), one showed only the 90-bp amplified band, and 4 showed both 165-bp and 90-bp bands. Both bands were seen more frequently in blast crisis than in remission and chronic phase.
采用RNA聚合酶链反应检测20例慢性髓性白血病（CML）患者的bcr/abl融合基因，该反应以mRNA为起始材料，通过逆转录酶生成cDNA，随后进行PCR扩增（RNA/PCR）。以所有20例CML患者（包括3例Ph（-）CML）的外周血细胞作为mRNA来源，成功扩增了跨越bcr/abl连接区的序列，CML患者来源的细胞系K562也用于实验。当使用3名正常个体的单核细胞、2例淋巴瘤患者的细胞以及细胞系HL-60时，未观察到扩增。融合mRNA中bcr外显子3的有无通过扩增片段的大小来确定。在20例CML患者中，15例仅显示165bp的扩增条带（表明保留bcr外显子3），1例仅显示90bp的扩增条带，4例同时显示165bp和90bp的条带。与缓解期和慢性期相比，急变期更常出现这两条条带。
