Detection of Philadelphia chromosome-positive cells from glass slide smears using the polymerase chain reaction.
Detection of Philadelphia chromosome-positive cells from glass slide smears using the polymerase chain reaction.
复制标题
使用聚合酶链式反应从载玻片涂片中检测费城染色体阳性细胞。
作者：
C. Hanson;E. Holbrook;S. Sheldon;B. Schnitzer;M. Roth
应用Southern和北方杂交及聚合酶链反应（PCR）技术对慢性粒细胞白血病（CML）bcr-abl基因复合体进行了分析。由于新鲜或冷冻保存的细胞可能并不总是可用于分子分析，我们研究了使用常规制备的血液或骨髓载玻片涂片作为细胞材料来源的可能性。使用改良的RNA提取程序直接从血液或骨髓涂片制备细胞RNA。从RNA合成cDNA，并使用bcr和ab 1特异性引物进行PCR扩增。应用此方法对21例CML患者中的21例进行了bcr-abl融合基因PCR检测。3例曾接受异基因骨髓移植（BMT）治疗CML的患者也进行了PCR研究。bcr-abl在1例患者中被短暂识别，在1例患者中在BMT后持续2年直至复发，在1例患者中在BMT后18个月内缺失。我们已经证明，PCR可以检测CML的bcr-abl基因使用材料从玻璃片涂片。这种技术可能是有用的，作为一种通用的方法，在评价档案血液标本的关键基因产物的表达。
Southern and Northern blot hybridization studies and the polymerase chain reaction (PCR) have been used to analyze the bcr-abl gene complex in chronic myelogenous leukemia (CML). Because fresh or cryopreserved cells may not always be available for molecular analyses, we investigated the possibility of using routinely prepared glass slide smears of blood or bone marrow as our source of cellular material. Cellular RNA was prepared directly from the blood or bone marrow smears using a modified RNA extraction procedure. cDNA was synthesized from RNA and amplified with PCR using bcr and abl-specific primers. Using this procedure, the bcr-abl fusion gene was detected by PCR in 21 of 21 patients with CML. Three patients who had undergone allogenic bone marrow transplantation (BMT) for CML were also studied by PCR. bcr-abl was identified transiently in one patient, persisted in one patient after BMT for 2 years until relapse occurred, and was absent in one patient to 18 months after BMT. We have shown that PCR can detect the bcr-abl gene of CML using material from glass-slide smears. This technique may be useful as a general approach in evaluating archival hematologic specimens for the expression of critical gene products.