The present invention relates to a method for detecting mycoplasma 16S rDNA 1.5 kilobase fragment amplified quantitative polymerase chain reaction (qPCR).rnrnThe method according to the invention is suitable for the detection of potentially any Prokaryote contamination, especially mycoplasma contamination, in cell cultures.rnrnAnother possible application can be its use in diagnosis and follow up of contamination in human, livestock, plants, food, or water for example.