检测遗传毒性的umu高通量微板试验
来源： 浏览量：497 更新日期：2010年1月22日
Environmental and Molecular Mutagenesis 43:10–19 (2004)
Use of a High-Throughput umu-Microplate Test System for Rapid Detection of Genotoxicity Produced by Mutagenic Carcinogens and Airborne Particulate Matter
Yoshimitsu Oda,1* Kunihiro Funasaka,2 Masaaki Kitano,2
Akihiko Nakama,2 and Taro Yoshikura2
1Osaka Prefectural Institute of Public Health, Osaka, Japan
2Osaka City Institute of Public Health and Environmental Sciences, Osaka, Japan
In the present study, we developed a rapid umumicroplate test system that uses the nitroreductaseand O-acetyltransferase-overproducing Salmonella typhimurium strain NM3009 and the O-acetyltransferase -overproducing S. typhimurium strain NM2009 to detect genotoxic activity in small volume samples. The assay was used to test the genotoxicity of several standard mutagens and environmental samples. Exponentially growing cultures of NM3009, NM2009, and the parental strain TA1535/pSK1002 were incubated in 96-well microplates with test chemicals both in the presence and in the absence of rat liver S9. The relative _-galactosidase activities were then determined colorimetrically using either chlorophenol red-_-D-galactopyranoside (CPRG) or O-nitrophenyl-_-D-galactopyranoside (ONPG) as a measure of umuC gene induction activity. The sensitivities of NM3009 without S9 mix and NM2009 with S9 mix to nitroarenes and aromatic amines were up to 24- to 75-fold higher than those of the parent strain. Induction of umuC gene expression was detected more readily with CPRG than ONPG. The umu-microplate assay also detected genotoxicity in organic extracts of particulate matter from air samples collected in Osaka City, Japan. The pattern of the responses suggested that the genotoxic activity of the particulate extract was due primarily to nitrated polycyclic aromatic hydrocarbons. Our results indicate that the umu-microplate assay may be a useful way of carrying out rapid screens for genotoxicity in small-volume environmental samples.