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

doi:10.1016/j.isci.2023.106724

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

iScience. 2023 Apr 26;26(5):106724. doi: 10.1016/j.isci.2023.106724. eCollection 2023 May 19.

Authors

Dominik Siegl¹, Marie Kruchem¹, Sandrine Jansky¹, Emma Eichler¹, Dorothe Thies¹, Udo Hartwig^{2

3}, Detlef Schuppan^{1

4

5

3}, Ernesto Bockamp^{1

4

3}

Affiliations

¹ Institute of Translational Immunology (TIM), University Medical Center of the Johannes Gutenberg-University, Mainz 55131, Germany.
² Department of Medicine III Hematology & Medical Oncology, University Medical Center of the Johannes Gutenberg-University, Mainz 55131, Germany.
³ Research Center for Immunotherapy, University Medical Center of the Johannes Gutenberg-University, Mainz 55131, Germany.
⁴ ImmuneNTech GmbH, Wendelsheim 55234, Germany.
⁵ Division of Gastroenterology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA 02215, USA.

Abstract

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.

Keywords: Microbiology; Mycology; Sequence analysis.