Department of Pathology and Laboratory Medicine, Dartmouth-Hitchcock Medical Center, Lebanon, New Hampshire, USA.
Dartmouth College, Hanover, New Hampshire, USA.
J Cutan Pathol. 2024 Feb;51(2):146-154. doi: 10.1111/cup.14540. Epub 2023 Oct 5.
While most melanocytic neoplasms can be classified as either benign or malignant by histopathology alone, ancillary molecular diagnostic tests can be necessary to establish the correct diagnosis in challenging cases. Currently, the detection of copy number variations (CNVs) by fluorescence in situ hybridization and chromosomal microarray (CMA) are the most popular methods, but remain expensive and inaccessible. We aim to develop a relatively inexpensive, fast, and accessible molecular assay to detect CNVs relevant to melanoma using droplet digital polymerase chain reaction (ddPCR) technology.
In this proof-of-concept study, we evaluated CNVs in MYC and MYB genes from 73 cases of benign nevi, borderline melanocytic lesions, and primary and metastatic melanoma at our institution from 2015 to 2022. A multiplexed ddPCR assay and CMA were performed on each sample, and the results were compared.
Concordance analysis of ddPCR with CMA for quantification of MYC and MYB CNVs revealed a sensitivity and specificity of 89% and 86% for MYC and 83% and 74% for MYB, respectively.
We demonstrate the first use of a multiplexed ddPCR assay to identify CNVs in melanocytic neoplasms. With further improvement and validation, ddPCR may represent a low-cost and rapid tool to aid in the diagnosis of histopathologically ambiguous melanocytic tumors.
虽然大多数黑色素细胞肿瘤仅通过组织病理学即可被归类为良性或恶性，但在具有挑战性的病例中，可能需要辅助的分子诊断测试来确定正确的诊断。目前，荧光原位杂交和染色体微阵列（CMA）的拷贝数变异（CNV）检测是最流行的方法，但仍然昂贵且难以获得。我们旨在开发一种相对廉价、快速且易于获得的分子检测方法，使用液滴数字聚合酶链反应（ddPCR）技术检测与黑色素瘤相关的 CNVs。
在这项概念验证研究中，我们评估了 2015 年至 2022 年期间我院 73 例良性痣、交界性黑色素细胞病变以及原发性和转移性黑色素瘤中 MYC 和 MYB 基因的 CNVs。对每个样本进行了多重 ddPCR 检测和 CMA，并比较了结果。
ddPCR 与 CMA 对 MYC 和 MYB CNVs 定量的一致性分析显示，MYC 的灵敏度和特异性分别为 89%和 86%，MYB 的灵敏度和特异性分别为 83%和 74%。
我们首次展示了使用多重 ddPCR 检测来识别黑色素细胞肿瘤中的 CNVs。通过进一步改进和验证，ddPCR 可能代表一种低成本、快速的工具，有助于诊断组织病理学上有疑问的黑色素瘤。
