A PCR protocol to establish standards for routine mycoplasma testing that by design detects over ninety percent of all known mycoplasma species

Mycoplasma infection leads to false and non-reproducible scientific data and poses a risk to human health. Despite strict guidelines calling for regular myco-plasma screening, there is no universal and widely established standard proced-ure. Here, we describe a reliable and cost-effective PCR method that establishes a universal protocol for mycoplasma testing. The applied strategy utilizes ultra -conserved eukaryotic and mycoplasma sequence primers covering by design 92% of all species in the six orders of the class Mollicutes within the phylum My-coplasmatota and is applicable to mammalian and many non-mammalian cell types. This method can stratify mycoplasma screening and is suitable as a com-mon standard for routine mycoplasma testing.