Electrochemical mutagen screening using microbial chip.
Electrochemical microbial chip for mutagen screening were microfabricated and characterized by scanning electrochemical microscopy (SECM). Salmonella typhimurium TA1535 with a plasmid pSK1002 carrying a umuC'-'lacZ fusion gene was used for the whole cell mutagen sensor. The TA1535/pSK1002 cells were exposed to mutagen solutions containing 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamido (AF-2), mitomycin C (MMC) or 2-aminoanthracene (2-AA) and embedded in a microcavity (5nl) on a glass substrate using collagen gel. The beta-galactosidase expression on the microbial chip was electrochemically monitored using p-aminophenyl-beta-d-galactopyranoside (PAPG) as the enzymatic substrate. This system has several advantages compared with the conventional umu test: drastic reduction of the sample volume, less time-consuming for beta-galactosidase detection (free from substrate reaction time) and lower detection limit for the three mutagens (AF-2, MMC, 2-AA). Finally, a multi-sample assay was carried out using the microbial array chip with four microcavities.
Matsui
Nobuto
N
Graduate School of Environmental Studies, Tohoku University, 6-6-11 Aramaki Aoba, Aoba-ku, Sendai 980-8579, Japan.
Kaya
Takatoshi
T
Nagamine
Kuniaki
K
Yasukawa
Tomoyuki
T
Shiku
Hitoshi
H
Matsue
Tomokazu
T
D001681:Biological Assay / Q000295:instrumentation* / Q000379:methods; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods; D004563:Electrochemistry / Q000295:instrumentation* / Q000379:methods; D004867:Equipment Design; D019544:Equipment Failure Analysis; D004926:Escherichia coli / Q000187:drug effects* / Q000201:enzymology*; D009153:Mutagens / Q000008:administration & dosage* / Q000032:analysis*; D015203:Reproducibility of Results; D012486:Salmonella typhimurium / Q000187:drug effects* / Q000201:enzymology; D012680:Sensitivity and Specificity; D001616:beta-Galactosidase / Q000032:analysis
D009153:Mutagens; D001616:beta-Galactosidase