New method for evaluation of genotoxicity, based on the use of real-time PCR and lysogenic gram-positive and gram-negative bacteria.
A method for the detection of the SOS response as measured by the liberation of resident prophages from the genomes of their hosts is described. It is based on the use of two converging oligonucleotides that flank the attP attachment site of the phage as primers for real-time PCR. Amplification was observed only after the phage DNA became excised. The system responds to both chemicals and physical conditions. Quantitative data on the concentration and/or potency of the genotoxic condition were obtained. Results can be achieved within 1 day and are less susceptible to possible toxic effects than phage generation or other methods that require DNA synthesis. The use of both gram-positive and gram-negative bacteria widens the range of compounds that can be tested because it eliminates impermeability problems derived from the particular composition of each cell wall type.
Soberón
Nora
N
Area de Microbiología, Facultad de Medicina, Universidad de Oviedo, Julián Clavería 6, 33006 Oviedo, Spain.
Martín
Rebeca
R
Suárez
Juan E
JE
D001287:Attachment Sites, Microbiological / Q000235:genetics; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics / Q000821:virology*; D007780:Lacticaseibacillus casei / Q000187:drug effects / Q000235:genetics / Q000821:virology*; D008242:Lysogeny; D016685:Mitomycin / Q000633:toxicity; D009152:Mutagenicity Tests / Q000379:methods*; D009153:Mutagens / Q000633:toxicity*; D009841:Oligonucleotides / Q000235:genetics; D016133:Polymerase Chain Reaction; D013014:SOS Response, Genetics / Q000187:drug effects / Q000235:genetics*; D014466:Ultraviolet Rays; D014775:Virus Activation / Q000187:drug effects / Q000235:genetics* / Q000528:radiation effects
D009153:Mutagens; D009841:Oligonucleotides; D016685:Mitomycin