Fast-track DRESSA: a bioassay for fast, sensitive, and selective detection of halogenated and polycyclic aromatic hydrocarbons

被引:22
作者
Kasai, A
Hiramatsu, N
Meng, YM
Yao, J
Maeda, S
Kitamura, M [1 ]
机构
[1] Univ Yamanashi, Interdisciplinary Grad Sch Med & Engn, Dept Mol Signaling, Tamaho, Yamanashi 4093898, Japan
[2] Univ Yamanashi, Interdisciplinary Grad Sch Med & Engn, Dept Biochem, Tamaho, Yamanashi 4093898, Japan
关键词
biosensor; bioassay; dioxin; dioxin-responsive element; secreted alkaline phosphatase;
D O I
10.1016/j.ab.2004.10.004
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:84 / 88
页数:5
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