A PCR protocol to establish standards for routine mycoplasma testing that by design detects over ninety percent of all known mycoplasma species
Dominik Siegl,
Marie Kruchem,
Sandrine Jansky,
Emma Eichler,
Dorothe Thies,
Udo Hartwig,
Detlef Schuppan,
Ernesto Bockamp
Affiliations
Dominik Siegl
Institute of Translational Immunology (TIM), University Medical Center of the Johannes Gutenberg-University, Mainz 55131, Germany; Corresponding author
Marie Kruchem
Institute of Translational Immunology (TIM), University Medical Center of the Johannes Gutenberg-University, Mainz 55131, Germany
Sandrine Jansky
Institute of Translational Immunology (TIM), University Medical Center of the Johannes Gutenberg-University, Mainz 55131, Germany
Emma Eichler
Institute of Translational Immunology (TIM), University Medical Center of the Johannes Gutenberg-University, Mainz 55131, Germany
Dorothe Thies
Institute of Translational Immunology (TIM), University Medical Center of the Johannes Gutenberg-University, Mainz 55131, Germany
Udo Hartwig
Department of Medicine III Hematology & Medical Oncology, University Medical Center of the Johannes Gutenberg-University, Mainz 55131, Germany; Research Center for Immunotherapy, University Medical Center of the Johannes Gutenberg-University, Mainz 55131, Germany
Detlef Schuppan
Institute of Translational Immunology (TIM), University Medical Center of the Johannes Gutenberg-University, Mainz 55131, Germany; ImmuneNTech GmbH, Wendelsheim 55234, Germany; Division of Gastroenterology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA 02215, USA; Research Center for Immunotherapy, University Medical Center of the Johannes Gutenberg-University, Mainz 55131, Germany
Ernesto Bockamp
Institute of Translational Immunology (TIM), University Medical Center of the Johannes Gutenberg-University, Mainz 55131, Germany; ImmuneNTech GmbH, Wendelsheim 55234, Germany; Research Center for Immunotherapy, University Medical Center of the Johannes Gutenberg-University, Mainz 55131, Germany
Summary: Mycoplasma infection leads to false and non-reproducible scientific data and poses a risk to human health. Despite strict guidelines calling for regular mycoplasma screening, there is no universal and widely established standard procedure. 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 Mycoplasmatota and is applicable to mammalian and many non-mammalian cell types. This method can stratify mycoplasma screening and is suitable as a common standard for routine mycoplasma testing.
