Fast-track DRESSA: a bioassay for fast, sensitive, and selective detection of halogenated and polycyclic aromatic hydrocarbons
Fast-track DRESSA: a bioassay for fast, sensitive, and selective detection of halogenated and polycyclic aromatic hydrocarbons
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DOI：
10.1016/j.ab.2004.10.004
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发表时间：
2005-02-01
影响因子：
2.9
通讯作者：
Kitamura, M
中科院分区：
文献类型：
--
作者：
Kasai, A;Hiramatsu, N;Kitamura, M
二恶英和相关化学品通过芳香烃受体（AhR）引起各种毒性和生物效应。我们最近报道了一个哺乳动物细胞为基础的生物检测系统（二恶英响应元件为基础的传感通过分泌型碱性磷酸酶; DRESSA），可以检测二恶英和二恶英类化学品的高灵敏度。在这份报告中，我们描述了一种先进的方法（指定为“快速跟踪DRESSA”），实现快速，选择性和灵敏的检测二恶英和其他有毒化合物。通过优化细胞数和血清浓度的检测条件，快速跟踪DRESSA能够在6 h内检测0.5 pM的2，3，7，8-四氯二苯并-p-二恶英。它还能够在6-16小时内检测10 pM 3-甲基胆蒽、100 pM苯并[a]芘和100 pM β-萘啶酮。通过与AhR拮抗剂α-萘酚酮组合，可以消除非特异性假阳性反应。快速DRESSA技术具有快速、简便、灵敏、特异等优点，有望用于环境样品中二恶英类化合物的高通量筛选。(C)2004年爱思唯尔公司All rights reserved.
Dioxin and related chemicals cause a variety of toxic and biological effects via the aryl hydrocarbon receptor (AhR). We recently reported a mammalian cell-based bioassay system (dioxin-responsive-element-based sensing via secreted alkaline phosphatase; DRESSA) that can detect dioxin and dioxin-like chemicals with high sensitivity. In this report, we describe an advanced method (designated "fast-track DRESSA") that achieves fast, selective, and sensitive detection of dioxin and other toxic compounds. By optimization of assay conditions on cell number and serum concentration, the fast-track DRESSA enabled detection of 0.5 pM 2,3,7,8-tetrachlorodibenzo-p-dioxin within 6 h. It also enabled detection of 10 pM 3-methylcholanthrene, 100 pM benzo[a]pyrene, and 100 pM beta-naphthoflavone within 6-16 h. By combination with the AhR antagonist alpha-naphthoflavone, nonspecific, false-positive responses could be eliminated. Because of its time-saving property and easiness, sensitiveness, and specificity, the fast-track DRESSA would be advantageous for high-throughput screening of dioxin and dioxin-like compounds in environmental samples. (C) 2004 Elsevier Inc. All rights reserved.