玻璃体液和房水标本的无细胞 DNA 提取用于诊断和监测脉络膜视网膜淋巴瘤。
Cell-Free DNA Extraction of Vitreous and Aqueous Humor Specimens for Diagnosis and Monitoring of Vitreoretinal Lymphoma.
机构信息
Department of Pathology, University of Michigan, Ann Arbor;
Department of Pathology, University of Michigan, Ann Arbor; Department of Ophthalmology and Visual Sciences, Kellogg Eye Center, University of Michigan, Ann Arbor; Department of Human Genetics, University of Michigan, Ann Arbor; Rogel Cancer Center, University of Michigan, Ann Arbor; Center of Computational Medicine and Bioinformatics, University of Michigan, Ann Arbor; Center for RNA Biomedicine, University of Michigan, Ann Arbor; A. Alfred Taubman Medical Research Institute, University of Michigan, Ann Arbor; Section of Ophthalmology, Surgery Service, Veterans Administration Ann Arbor Healthcare System.
出版信息
J Vis Exp. 2024 Jan 12(203). doi: 10.3791/65708.
Vitreoretinal lymphoma (VRL) represents an aggressive lymphoma, often categorized as primary central nervous system diffuse large B-cell lymphoma. To diagnose VRL, specimens such as vitreous humor and, more recently, aqueous humor are collected. Diagnostic testing for VRL on these specimens includes cytology, flow cytometry, and molecular testing. However, both cytopathology and flow cytometry, along with molecular testing using cellular DNA, necessitate intact whole cells. The challenge lies in the fact that vitreous and aqueous humor typically have low cellularity, and many cells get destroyed during collection, storage, and processing. Moreover, these specimens pose additional difficulties for molecular testing due to the high viscosity of vitreous humor and the low volume of both vitreous and aqueous humor. This study proposes a method for extracting cell-free DNA from vitreous and aqueous specimens. This approach complements the extraction of cellular DNA or allows the cellular component of these specimens to be utilized for other diagnostic methods, including cytology and flow cytometry.
玻璃体内视网膜淋巴瘤 (Vitreoretinal lymphoma, VRL) 是一种侵袭性淋巴瘤，通常归类为原发性中枢神经系统弥漫性大 B 细胞淋巴瘤。为了诊断 VRL，采集玻璃体和最近的房水等标本。对这些标本进行 VRL 的诊断检测包括细胞学、流式细胞术和分子检测。然而，细胞病理学和流式细胞术以及使用细胞 DNA 的分子检测都需要完整的整个细胞。挑战在于玻璃体和房水通常细胞数量较少，并且在采集、储存和处理过程中许多细胞会被破坏。此外，由于玻璃体液的高粘度以及玻璃体和房水的体积均较低，这些标本给分子检测带来了额外的困难。本研究提出了一种从玻璃体液和房水标本中提取无细胞 DNA 的方法。这种方法可以补充细胞内 DNA 的提取，或者允许这些标本的细胞成分用于其他诊断方法，包括细胞学和流式细胞术。
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