Highly sensitive reporter gene assay for dioxins in human plasma by using cycloheximide as an enhancer substance.
A reporter gene assay (RGA) that uses a mouse liver recombinant Hepa1c1c7 containing the firefly luciferase gene was developed to screen for dioxins in human plasma. For a high-sensitivity method, the addition of cycloheximide to the culture medium brought about a fivefold increase in the sensitivity. The detection limit was 0.1 pg/microL/well. Aryl hydrocarbon receptor (AhR) binding affinity factors (AhR-BAF), calculated from the effect concentration 50 (EC(50)) value, showed approximately the same values as those in WHO-TEF (2006). A significant correlation between RGA and the conventional gas chromatography/mass spectrometry (GC/MS) method was obtained.
Saito
Koichi
K
Department of Analytical Chemistry, Faculty of Pharmaceutical Sciences, Hoshi University, Shinagawa, Tokyo, Japan. k-saito@hoshi.ac.jp
Matsunaga
Haruyuki
H
Ohmura
Atsuko
A
Takekuma
Mikiko
M
Matsuki
Yasuhiko
Y
Nakazawa
Hiroyuki
H
D000818:Animals; D045744:Cell Line, Tumor; D003513:Cycloheximide; D004147:Dioxins / Q000097:blood*; D017930:Genes, Reporter; D006801:Humans; D007202:Indicators and Reagents; D008099:Liver / Q000378:metabolism; D049409:Luciferases, Firefly / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D018336:Receptors, Aryl Hydrocarbon / Q000378:metabolism
D004147:Dioxins; D007202:Indicators and Reagents; D018336:Receptors, Aryl Hydrocarbon; D003513:Cycloheximide; D049409:Luciferases, Firefly