Yamashita S, Umemura T, Sadamura S, Yufu Y, Nishimura J, Nawata H
Third Department of Internal Medicine, Faculty of Medicine, Kyushu University, Japan.
Leukemia. 1994 Aug;8(8):1409-10.
We describe the polymerase chain reaction (PCR) method using stained bone marrow smears as sources of RNA. The amount of extractable RNA decreased during the process of making and staining bone marrow smears. The sensitivity of the reverse transcriptase-based polymerase chain reaction (RT-PCR) method for detecting target mRNA-positive cells in 5 x 10(5) suspended cells and stained bone marrow smears were 1:10(5) and 1:5000, when we used K562 cells. The bone marrow smears of 21 patients with chronic myelogenous leukemia (CML) were examined using this method. We extracted RNA from stained specimens stored at room temperature for 5-14 years. Twelve of 21 (57%) smears showed positive results for bcr/abl. The carrier RNA improved the recovery when added at the step of RNA extraction. These data indicate that mRNA is present in stained bone marrow smears for at least 14 years and that the sensitivity of RT-PCR is adequate for molecular analysis.
我们描述了一种以经染色的骨髓涂片作为RNA来源的聚合酶链反应（PCR）方法。在制作和染色骨髓涂片的过程中，可提取RNA的量会减少。当我们使用K562细胞时，基于逆转录酶的聚合酶链反应（RT-PCR）方法在检测5×10⁵个悬浮细胞和经染色的骨髓涂片中靶mRNA阳性细胞时的灵敏度分别为1:10⁵和1:5000。我们使用该方法检查了21例慢性髓性白血病（CML）患者的骨髓涂片。我们从室温下保存5至14年的染色标本中提取RNA。21份涂片中的12份（57%）bcr/abl检测呈阳性。在RNA提取步骤中添加载体RNA可提高回收率。这些数据表明，mRNA在经染色的骨髓涂片中至少存在14年，并且RT-PCR的灵敏度足以进行分子分析。
