Fast-track DRESSA: a bioassay for fast, sensitive, and selective detection of halogenated and polycyclic aromatic hydrocarbons.
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.
Kasai
Ayumi
A
Department of Biochemistry, Interdisciplinary Graduate School of Medicine and Engineering, University of Yamanashi, Tamaho, Yamanashi 409-3898, Japan.
Hiramatsu
Nobuhiko
N
Meng
Yiman
Y
Yao
Jian
J
Maeda
Shuichiro
S
Kitamura
Masanori
M
D000469:Alkaline Phosphatase; D000818:Animals; D001564:Benzo(a)pyrene / Q000032:analysis; D045744:Cell Line, Tumor; D004147:Dioxins / Q000032:analysis; D006846:Hydrocarbons, Halogenated / Q000032:analysis*; D008748:Methylcholanthrene / Q000032:analysis; D051379:Mice; D000072317:Polychlorinated Dibenzodioxins / Q000032:analysis; D011084:Polycyclic Aromatic Hydrocarbons / Q000032:analysis*; D015203:Reproducibility of Results; D013997:Time Factors; D019324:beta-Naphthoflavone / Q000032:analysis
D004147:Dioxins; D006846:Hydrocarbons, Halogenated; D000072317:Polychlorinated Dibenzodioxins; D011084:Polycyclic Aromatic Hydrocarbons; D001564:Benzo(a)pyrene; D008748:Methylcholanthrene; D019324:beta-Naphthoflavone; D000469:Alkaline Phosphatase