摘要
应用改良的一步法RNA提取方法分离慢性粒细胞白血病(慢粒)患者外周血或骨髓标本总RNA,以慢粒急性红白血病变细胞株为阳性对照,急件早幼粒细胞性白血病细胞株为阴性对照,建立逆转录/套式聚合酶链反应(RT,套式PCR)检测断裂点簇集区一abl融合基因。第一次PCR的灵敏度为1:10 ̄4水平,第二次PCR后灵敏度提高至1:10 ̄5水平。特异性扩增产物经用地高辛素标记寡核苷酸探针Southern杂交得到证实。
sing the modified single一step method to extract RNA from the peripheral blood or bone marrow of the patients with chronic mveloid leukemia. the author established RT/PCR and nested PCR to de-tect bcr一abl fused gene with the positive control K562 cell line and negative control HL一60 cell lineThe sensitivity of the first round of PCR is 1:10 ̄4 and it increases to 1:10 ̄5 after the second round ofPCR The special products have been confirmed by the Southern blot hybridization with Digoxigene la-beled oligonucleotide probe.