帕金森病患者下颌下腺α-突触核蛋白实时液滴震动诱导转化。
α-Synuclein real-time quaking-induced conversion in the submandibular glands of Parkinson's disease patients.
机构信息
Department of Biomedical Sciences, Parkinson's Disorder Research Program, Iowa Center for Advanced Neurotoxicology, Iowa State University, Ames, Iowa, USA.
Department of Neurology and Pharmacology, Neurosurgery, Radiology, and Kinesiology, Penn State Milton S. Hershey Medical Center, Hershey, Pennsylvania, USA.
出版信息
Mov Disord. 2020 Feb;35(2):268-278. doi: 10.1002/mds.27907. Epub 2019 Nov 23.
BACKGROUND
Identification of a peripheral biomarker is a major roadblock in the diagnosis of PD. Immunohistological identification of p-serine 129 α-synuclein in the submandibular gland tissues of PD patients has been recently reported.
OBJECTIVE
We report on a proof-of-principle study for using an ultra-sensitive and specific, real-time quaking-induced conversion assay to detect pathological α-synuclein in the submandibular gland tissues of PD patients.
METHODS
The α-synuclein real-time quaking-induced conversion assay was used to detect and quantify pathological α-synuclein levels in PD, incidental Lewy body disease, and control submandibular gland tissues as well as in formalin-fixed paraffin-embedded sections.
RESULTS
We determined the quantitative seeding kinetics of pathological α-synuclein present in submandibular gland tissues from autopsied subjects using the α-synuclein real-time quaking-induced conversion assay. A total of 32 cases comprising 13 PD, 3 incidental Lewy body disease, and 16 controls showed 100% sensitivity and 94% specificity. Interestingly, both PD and incidental Lewy body disease tissues showed 100% concordance for elevated levels of pathological α-synuclein seeding activity compared to control tissues. End-point dilution kinetic analyses revealed that the submandibular gland had a wide dynamic range of pathological α-synuclein seeding activity.
CONCLUSIONS
Our results are the first to demonstrate the utility of using the real-time quaking-induced conversion assay on peripherally accessible submandibular gland tissues and formalin-fixed paraffin-embedded tissue sections to detect PD-related pathological changes with high sensitivity and specificity. Additionally, the detection of seeding activity from incidental Lewy body disease cases containing immunohistochemically undetected pathological α-synuclein demonstrates the α-synuclein real-time quaking-induced conversion assay's potential utility for identifying prodromal PD in submandibular gland tissues. © 2019 International Parkinson and Movement Disorder Society.
背景
鉴定外周生物标志物是 PD 诊断的主要障碍。最近有报道称，在 PD 患者的颌下腺组织中免疫组织化学鉴定 p-丝氨酸 129 α-突触核蛋白。
目的
我们报告了一项原理验证研究，使用超灵敏和特异的实时液滴式数字 PCR 检测 PD 患者颌下腺组织中的病理性 α-突触核蛋白。
方法
使用 α-突触核蛋白实时液滴式数字 PCR 检测和定量 PD、偶然 Lewy 体病和对照颌下腺组织以及福尔马林固定石蜡包埋切片中的病理性 α-突触核蛋白水平。
结果
我们使用 α-突触核蛋白实时液滴式数字 PCR 确定了从尸检对象颌下腺组织中存在的病理性 α-突触核蛋白的定量接种动力学。共 32 例，包括 13 例 PD、3 例偶然 Lewy 体病和 16 例对照，具有 100%的敏感性和 94%的特异性。有趣的是，与对照组织相比，PD 和偶然 Lewy 体病组织中的病理性 α-突触核蛋白接种活性均显示出 100%的一致性升高。终点稀释动力学分析表明，颌下腺具有广泛的病理性 α-突触核蛋白接种活性动态范围。
结论
我们的研究结果首次证明了使用实时液滴式数字 PCR 检测外周可及的颌下腺组织和福尔马林固定石蜡包埋组织切片中 PD 相关病理性改变的实用性，具有高灵敏度和特异性。此外，从含有免疫组织化学未检测到病理性 α-突触核蛋白的偶然 Lewy 体病病例中检测到的接种活性，证明了实时液滴式数字 PCR 检测 α-突触核蛋白在颌下腺组织中识别前驱期 PD 的潜在用途。
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