Novel mutation assay with high sensitivity based on direct measurement of genomic DNA alterations: comparable results to the Ames test.
Almost all of the methodologies developed to date to assay the potential mutagenicity of chemical substances are based on detection of altered phenotypic traits. The alternative approach of directly screening the whole genome for mutations is not feasible because of the logistics of carrying out mass sequencing of genes. Here we describe a novel and highly sensitive mutation assay, which we term the 'genome profiling-based mutation assay' (GPMA) that directly detects mutations generated in genomic DNA. We used GPMA to detect mutations caused by known mutagens such as AF2 and ethidium bromide even at concentrations of 30 ppb. The number of mutations detected was dependent on the number of generations in culture and the concentrations of the mutagens. Almost complete agreement was observed between GPMA and the Ames test in the discrimination of mutagens (63 out of 64). Owing to the high sensitivity of GPMA, the effects of long-term and low-dose exposures and the influence of chemicals of low solubility can also be screened. Thus, genotype-based GPMA can complement the Ames test, which is the standard technology in this field and is based on phenotypic traits.
Futakami
Masae
M
Department of Functional Materials Science, Saitama University, 255 Shimo-Okubo, Sakura-ku, Saitama 338-8570, Japan.
Salimullah
Md
M
Miura
Takashi
T
Tokita
Sumio
S
Nishigaki
Koichi
K
D004247:DNA / Q000737:chemistry*; D004252:DNA Mutational Analysis / Q000379:methods*; D004269:DNA, Bacterial / Q000737:chemistry / Q000187:drug effects; D048168:Escherichia coli K12 / Q000187:drug effects / Q000235:genetics; D004996:Ethidium / Q000494:pharmacology; D005668:Furylfuramide / Q000494:pharmacology; D009152:Mutagenicity Tests / Q000379:methods*; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity
D004269:DNA, Bacterial; D005668:Furylfuramide; D004247:DNA; D004996:Ethidium