Laboratory of Immunology, National Eye Institute, National Institutes of Health, Bethesda, Maryland.
Office of the Director, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Rockville, Maryland.
Am J Ophthalmol. 2014 Mar;157(3):728-34. doi: 10.1016/j.ajo.2013.12.014. Epub 2013 Dec 15.
To use micro-ribonucleic acid (microRNA) profiles in the vitreous for differential diagnosis of primary vitreoretinal lymphoma and uveitis.
Prospective cross-sectional study.
This prospective cross-sectional study included 17 diffuse large B-cell primary vitreoretinal lymphoma and 12 uveitis patients. The supernatant of ocular fluid was subjected to total RNA extraction, followed by complementary deoxyribonucleic acid (cDNA) synthesis. Selected samples (primary vitreoretinal lymphoma, n = 3; uveitis, n = 3) were arrayed by a real-time polymerase chain reaction (RT-PCR)-based microRNA panel that detects 168 human mature microRNAs. The markers promising in distinct levels between uveitis and lymphoma were further tested for in all the other 23 samples by individual RT-PCR analysis.
Of 168 microRNAs in the array, 66.5% were detectable with consistent higher microRNA-484, microRNA-197, and microRNA-132 in the primary vitreoretinal lymphoma vitreous and higher microRNA-155, microRNA-200c, and microRNA-22* in the uveitic ocular fluids. The results were normalized by different combinations of 7 control microRNAs (microRNA-103, microRNA-191, microRNA-42-5p, microRNA-16, microRNA-425, microRNA-93, and microRNA-451). After optimization, normalization against microRNA-16 was equally as reliable as the average of the 7 control microRNAs. Individual assays of all samples supported the pattern yielded from the array analysis. But only microRNA-155 was significantly higher in the uveitic vitreous compared to that with lymphoma.
Mature microRNAs are detectable in ocular fluid samples. Primary vitreoretinal B-cell lymphoma and uveitis might be characterized by distinct microRNA signatures. Quantification of ocular microRNA-155 might be helpful in the differential diagnosis of these 2 diseases.
利用玻璃体中的 microRNA 谱对原发性玻璃体视网膜淋巴瘤和葡萄膜炎进行鉴别诊断。
前瞻性横断面研究。
本前瞻性横断面研究纳入了 17 例弥漫性大 B 细胞原发性玻璃体视网膜淋巴瘤和 12 例葡萄膜炎患者。对眼液上清液进行总 RNA 提取，然后进行互补脱氧核糖核酸（cDNA）合成。选择部分样本（原发性玻璃体视网膜淋巴瘤，n=3；葡萄膜炎，n=3）进行实时聚合酶链反应（RT-PCR）为基础的 microRNA 阵列检测，该阵列检测 168 个人类成熟 microRNA。在其他 23 例样本中，通过单独的 RT-PCR 分析进一步检测在淋巴瘤和葡萄膜炎之间有明显差异水平的标记物。
在该阵列的 168 个 microRNA 中，66.5%可检测到玻璃体中 microRNA-484、microRNA-197 和 microRNA-132 水平较高，而在葡萄膜炎眼液中则可检测到 microRNA-155、microRNA-200c 和 microRNA-22*水平较高。通过 7 个对照 microRNA（microRNA-103、microRNA-191、microRNA-42-5p、microRNA-16、microRNA-425、microRNA-93 和 microRNA-451）的不同组合进行归一化。经过优化，microRNA-16 的归一化与 7 个对照 microRNA 的平均值一样可靠。所有样本的单独检测结果均支持阵列分析产生的模式。但只有 microRNA-155 在葡萄膜炎玻璃体中显著高于淋巴瘤。
成熟的 microRNA 可在眼液样本中检测到。原发性玻璃体视网膜 B 细胞淋巴瘤和葡萄膜炎可能具有不同的 microRNA 特征。定量检测眼液中的 microRNA-155 可能有助于这两种疾病的鉴别诊断。
