Potential Diagnosis of Vitreoretinal Lymphoma by Detection of MYD88 Mutation in Aqueous Humor With Ultrasensitive Droplet Digital Polymerase Chain Reaction
Potential Diagnosis of Vitreoretinal Lymphoma by Detection of MYD88 Mutation in Aqueous Humor With Ultrasensitive Droplet Digital Polymerase Chain Reaction
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DOI：
10.1001/jamaophthalmol.2018.2887
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发表时间：
2018-10-01
影响因子：
8.1
通讯作者：
Kuiper, Jonas J. W.
中科院分区：
文献类型：
--
作者：
Hiemcke-Jiwa, Laura S.;Ten Darn-van Loon, Ninette H.;Kuiper, Jonas J. W.
重要提示对怀疑患有玻璃体视网膜淋巴瘤（VRL）的患者进行诊断检查主要基于玻璃体液分析，包括最近出现的髓样分化原发反应基因88（MYD 88）突变分析。房水穿刺术是一种相对侵入性较小且比玻璃体液标本采集更安全的方法，目的探讨高灵敏度液滴数字聚合酶链反应（ddPCR）检测MYD 88 L265 P基因突变的可行性。在VRL患者的玻璃体液和房水中是可行的。这项队列研究包括2005年8月至2017年8月在荷兰乌得勒支大学医学中心接受治疗的VRL患者的房水和玻璃体液样本。在MYD 88 L265 P分析之前，使用内部验证的ddPCR平台对眼液进行随机化和设盲。主要结果和指标采用ddPCR检测AH和VF中MYD 88 L265 P突变的存在。结果本研究包括来自63个个体的96个样本，其中包括23例VRL患者（其中10名女性和13名男性，平均[SD]年龄为72 [7.3]岁）和40名葡萄膜炎患者（其中23名女性和17名男性，平均[SD]年龄为58 [20.9]岁）。在23例VRL患者中的17例（74%）中，检测到MYD 88 L265 P;在任何葡萄膜炎患者中均未检测到MYD 88 L265 P。在玻璃体液和眼房水样本中均可检测到。在配对样本中，在MYD 88 L265 P阳性玻璃体液样本的9个房水样本中的8个（89%）中检测到突变。在玻璃体液中，MYD 88 ddPCR检测显示灵敏度为75%（95% CI，50%-92%），阳性预测值为100%;在房水中，灵敏度为67%（95% CI，42%-92%），阳性预测值为100%。两种液体的特异性均为100%。治疗后，突变不再检测到任何眼fluids.CONCLUSIONS和RELEVANCE之间的高度一致性的房水和玻璃体液样本表明，使用的容易接近的房水是几乎一样的信息作为玻璃体液中的关键体细胞突变的识别VRL患者。这种方法可以提供额外的微创工具，用于准确诊断、复发检测和治疗监测。
IMPORTANCE The diagnostic workup of patients suspected of having vitreoretinal lymphoma (VRL) is primarily based on vitreous fluid analysis, including the recently emerging myeloid differentiation primary response gene 88 (MYD88) mutation analysis. Aqueous humor paracentesis is a relatively less invasive and safer procedure than taking vitreous fluid specimens, and aqueous humor-based MYD88 mutation analysis would provide an additional liquid biopsy tool to diagnose and monitor patients with VRL.OBJECTIVE To investigate whether the detection of MYD88 L265P by highly sensitive droplet digital polymerase chain reaction (ddPCR) is feasible in the vitreous fluid and aqueous humor of patients with VRL.DESIGN, SETTING. AND PARTICIPANTS This cohort study includes aqueous humor and vitreous fluid samples from patients with VRL who were treated at the University Medical Center Utrecht, in Utrecht, the Netherlands, from August 2005 to August 2017 Ocular fluids were randomized and masked before MYD88 L265P analysis, which was performed using an in-house validated ddPCR platform. Patients with uveitis were included as a comparison group.MAIN OUTCOMES AND MEASURES The presence of MYD88 L265P mutation detected by ddPCR in AH and VF.RESULTS The study included 96 samples from 63 individuals, including 23 patients with VRL (of whom 10 were female and 13 male, with a mean [SD] age of 72 [7.3] years) and 40 individuals with uveitis (of whom 23 were female and 17 male, with a mean [SD] age of 58 [20.9] years). In 17 of 23 patients with VRL (74%), MYD88 L265P was detected; it was not detected in any of the patients with uveitis. It was detectable in both vitreous fluid and aqueous humor samples. In the paired samples, the mutation was detected in 8 of 9 aqueous humor samples (89%) of the MYD88 L265P-positive vitreous fluid samples. In vitreous fluid, the MYD88 ddPCR test showed a sensitivity of 75% (95% CI, 50%-92%) and a positive predictive value of 100%; in aqueous humor, sensitivity was 67% (95% CI, 42%-92%), and positive predictive value was 100%. Specificity was 100% in both fluids. After treatment, the mutation was no longer detectable in any ocular fluids.CONCLUSIONS AND RELEVANCE The high concordance between aqueous humor and vitreous fluid samples suggests that use of the easily accessible aqueous humor is nearly as informative as vitreous fluid in the identification of key somatic mutations in patients with VRL. This approach may provide an additional minimally invasive tool for accurate diagnosis, detection of recurrence, and monitoring of treatment.