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            引用本文:   劉海生, 袁文理, 陳永鈺, 陳晨, 田野, 顏曉梅. 尿液細胞外囊泡的單顆粒水平多參數定量表征. 分析化學, 2020, 48(10): 1305-1314. doi:  10.19756/j.issn.0253-3820.201443 [復制]

            Citation:   LIU Hai-Sheng , YUAN Wen-Li , CHEN Yong-Yu , CHEN Chen , TIAN Ye , YAN Xiao-Mei . Multi-parameter and Quantitative Characterization of Urinary Extracellular Vesicles at Single-Particle Level. Chinese Journal of Analytical Chemistry, 2020, 48(10): 1305-1314. doi: 10.19756/j.issn.0253-3820.201443 [復制]

            尿液細胞外囊泡的單顆粒水平多參數定量表征

            通訊作者:  顏曉梅, xmyan@xmu.edu.cn

            收稿日期: 2020-07-29

            基金項目: 本文系國家自然科學基金項目(Nos.21934004,21627811)和國家基礎科學人才培養基金項目(No.J1310024)資助

            Multi-parameter and Quantitative Characterization of Urinary Extracellular Vesicles at Single-Particle Level

            Corresponding author:  YAN Xiao-Mei , xmyan@xmu.edu.cn

            Received Date:  2020-07-29

            Fund Project:  This work was supported by the National Natural Science Foundation of China (Nos. 21934004, 21627811) and the National Fund for Fostering Talents of Basic Science (No. J1310024).

            尿液來源細胞外囊泡(Urinary extracellular vesicles,uEVs)作為新一類液體活檢標志物,在疾病診斷、治療監控領域具有廣闊的應用前景。然而,由于缺乏相應的表征技術,研究者對于納米尺度uEVs的粒徑分布、顆粒濃度以及生化表型等均缺乏了解。EVs具有高度個體差異性和多樣性,因此迫切需要發展高通量、多參數的單顆粒檢測技術?;诒狙芯拷M前期工作中結合瑞利散射和鞘流單分子熒光檢測技術研制的納米流式檢測裝置(Nano-flow cytometer,nFCM),本研究對超速離心法純化得到的uEVs的純度、濃度、粒徑、DNA、RNA和蛋白標志物在單顆粒水平進行多參數定量表征,其中EVs的檢測限低至40 nm。結果表明,uEVs的純度高達92%,10位健康受試者uEVs的顆粒濃度主要分布在3.0×109~8.9×1010個/mL之間,粒徑主要分布在40~120 nm之間。DNA和RNA主要位于較大粒徑的uEVs的內部,且顆粒之間的豐度存在很大的異質性。此外,使用nFCM對蛋白標志物CD9、CD63、CD81和CD24的陽性比率進行了測定。納米流式檢測技術可為uEVs的生化性質研究和臨床應用提供重要的技術支撐。

            關鍵詞:   尿液細胞外囊泡, 納米流式檢測技術, 純度, 粒徑分布, 核酸, 蛋白標志物
            Key words:   Urinary extracellular vesicles, Nano-flow cytometry, Purity, Size distribution, Nucleic acid, Protein marker
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            目錄

            尿液細胞外囊泡的單顆粒水平多參數定量表征

            劉海生, 袁文理, 陳永鈺, 陳晨, 田野, 顏曉梅

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