Oravakandy Manogna Das, Ahmed Faiq, Khera Rachna, Mundada Manasi, Murthy Sudha S, Rajappa Senthil J, Mohan M V T Krishna, Kumar B Pavan, Santa A
Department of Pathology and Laboratory Medicine, Basavatarakam Indo-American Cancer Hospital and Research Institute, Banjara Hills, Hyderabad, Telangana India.
Department of Laboratory Medicine, Gimcare Hospital, Kannur, 670007 Kerala India.
Indian J Hematol Blood Transfus. 2022 Oct;38(4):643-648. doi: 10.1007/s12288-022-01518-0. Epub 2022 Feb 1.
Nucleophosmin (NPM1) mutation is one of the most common recurring genetic abnormalities seen in acute myeloid leukemia (AML). Immunohistochemistry serves as a cost effective and simple surrogate testing method for detection of NPM1 mutation. This study was conducted to evaluate the frequency of aberrant cytoplasmic nucleophosmin 1 expression in leukemic blast cells on formalin fixed bone marrow trephine biopsy (BMB) sections and also to correlate this data with the reference molecular method (reverse transcriptase-polymerase chain reaction; RT-PCR and gene sequencing), where available. Immunostains were performed using mouse anti-NPM1 monoclonal antibody on 71 paraffin embedded bone marrow biopsies (BMB) of patients with AML of any French-American-British (FAB) subtype. Results of immunohistochemistry (IHC) were then compared with the reference molecular method. The proportion of NPM1 expression by immunostaining in AML cases was found to be 17%. Twelve of the total 71 cases demonstrated cytoplasmic nucleophosmin (NPMc+) on immunostaining. Eleven of the positive cases that were correlated with the molecular standard demonstrated mutation in exon 12 of NPM1 gene. Cytoplasmic nucleophosmin expression by immunostaining was found to be in complete agreement with the standard molecular method. In a resource restricted setup, the information from this study might help in providing an inexpensive and accurate detection method to facilitate introduction of this marker in diagnostic and prognostic workup of AML especially in patients showing normal karyotype and no common recurrent translocations.
核磷蛋白(NPM1)突变是急性髓系白血病(AML)中最常见的复发性基因异常之一。免疫组织化学是检测NPM1突变的一种经济有效且简单的替代检测方法。本研究旨在评估福尔马林固定骨髓活检(BMB)切片上白血病原始细胞中异常细胞质核磷蛋白1表达的频率,并在可行的情况下将该数据与参考分子方法(逆转录聚合酶链反应;RT-PCR和基因测序)相关联。使用小鼠抗NPM1单克隆抗体对71例任何法美英(FAB)亚型AML患者的石蜡包埋骨髓活检(BMB)进行免疫染色。然后将免疫组织化学(IHC)结果与参考分子方法进行比较。发现AML病例中通过免疫染色检测到的NPM1表达比例为17%。71例病例中有12例在免疫染色时显示细胞质核磷蛋白(NPMc+)。与分子标准相关的11例阳性病例在NPM1基因第12外显子中显示突变。发现通过免疫染色检测到的细胞质核磷蛋白表达与标准分子方法完全一致。在资源有限的情况下,本研究的信息可能有助于提供一种廉价且准确的检测方法,以便在AML的诊断和预后评估中引入该标志物,特别是在核型正常且无常见复发性易位的患者中。