Orazi A, Cotton J, Cattoretti G, Kotylo P K, John K, Manning J T, Neiman R S
Department of Pathology, Indiana University School of Medicine, Indianapolis 46202.
Am J Clin Pathol. 1994 Nov;102(5):640-5. doi: 10.1093/ajcp/102.5.640.
Terminal deoxynucleotidyl transferase (TdT) is a nuclear protein widely used as a marker for the diagnosis and classification of acute leukemia. The usual methods for detecting TdT require smears, imprints, or cryostat sections of unfixed tissue. A polyclonal rabbit anti-TdT serum was used to immunostain 54 routinely processed bone marrow sections from patients with acute leukemic disorders, using a recently described antigen-unmasking technique based on microwave oven heating. The specificity of this method of TdT analysis was confirmed by comparing the results obtained with conventional TdT analysis by indirect immunofluorescence. Terminal deoxynucleotidyl transferase reactivity was also evaluated in 44 nonmalignant and normal bone marrow specimens. All cases that were TdT-positive by immunofluorescence (41 of 42 "pre-B" and T-cell acute lymphoblastic leukemia, 2 of 5 acute myeloid leukemia, and 1 of 5 chronic myeloid leukemia in blast crisis) were also positive in paraffin sections. The percentage fluorescence positivity correlated with the percentage of immunoperoxidase stained cells in 44 of 45 cases. The remaining nonneoplastic and normal bone marrow biopsy specimens were TdT-negative. These results show that TdT immunoperoxidase staining of conventionally processed bone marrow specimens can be readily achieved by the use of a simple antigen-unmasking technique and may provide useful diagnostic information particularly in cases in which fresh tissue samples are unavailable.
末端脱氧核苷酸转移酶(TdT)是一种核蛋白,广泛用作急性白血病诊断和分类的标志物。检测TdT的常用方法需要未固定组织的涂片、印片或冰冻切片。使用一种基于微波炉加热的新描述的抗原暴露技术,用多克隆兔抗TdT血清对54例急性白血病患者常规处理的骨髓切片进行免疫染色。通过将该TdT分析方法的结果与间接免疫荧光法的传统TdT分析结果进行比较,证实了该方法的特异性。还对44例非恶性和正常骨髓标本进行了末端脱氧核苷酸转移酶反应性评估。所有免疫荧光法检测TdT阳性的病例(42例“前B”和T细胞急性淋巴细胞白血病中的41例、5例急性髓细胞白血病中的2例以及5例慢性髓细胞白血病急变期中的1例)在石蜡切片中也呈阳性。45例中有44例荧光阳性百分比与免疫过氧化物酶染色细胞百分比相关。其余非肿瘤性和正常骨髓活检标本TdT阴性。这些结果表明,通过使用简单的抗原暴露技术,可以很容易地对常规处理的骨髓标本进行TdT免疫过氧化物酶染色,并且可能提供有用的诊断信息,特别是在无法获得新鲜组织样本的情况下。