A Rapid and Sensitive Nucleic Acid Amplification Technique for Mycoplasma Screening of Cell Therapy Products

Lisa Dreolini; Mark Cullen; Eric Yung; Lawrence Laird; John R Webb; Brad H Nelson; Kevin A Hay; Miruna Balasundaram; Natasha Kekre; Robert A Holt

doi:10.1016/j.omtm.2020.01.009

A Rapid and Sensitive Nucleic Acid Amplification Technique for Mycoplasma Screening of Cell Therapy Products

Mol Ther Methods Clin Dev. 2020 Jan 30:17:393-399. doi: 10.1016/j.omtm.2020.01.009. eCollection 2020 Jun 12.

Authors

Lisa Dreolini¹, Mark Cullen¹, Eric Yung¹, Lawrence Laird¹, John R Webb^{2

3}, Brad H Nelson^{2

3

4}, Kevin A Hay^{5

6}, Miruna Balasundaram¹, Natasha Kekre⁷, Robert A Holt^{1

4

8}

Affiliations

¹ Canada's Michael Smith Genome Sciences Centre, BC Cancer, Vancouver, BC V5Z 1L3, Canada.
² Trev and Joyce Deeley Research Centre, BC Cancer, Victoria, BC V8R 6V5, Canada.
³ Department of Biochemistry and Microbiology, University of Victoria, Victoria, BC V8W 2Y2, Canada.
⁴ Department of Medical Genetics, University of British Columbia, Vancouver, BC V6T 1Z4, Canada.
⁵ Terry Fox Laboratory, BC Cancer, Vancouver, BC V5Z 1L3, Canada.
⁶ Department of Medicine, University of British Columbia, Vancouver, BC V6T 1Z3, Canada.
⁷ Blood and Marrow Transplant Program, Ottawa Hospital Research Institute, Ottawa, ON K1H 8L6, Canada.
⁸ Molecular Biology & Biochemistry, Simon Fraser University, Burnaby, BC V5A 1S6, Canada.

Abstract

Mycoplasma species (spp.) bacteria can infect cell cultures, posing a potential threat to recipients of cell therapy products. Conventional Mycoplasma testing methods are highly sensitive but typically require a minimum of 28 days to produce results. This delay is problematic if rapid results are needed to inform treatment decisions. Nucleic acid amplification technique (NAT) methods have been gaining favor for Mycoplasma testing due to their speed and specificity; however, they must first be qualified as meeting or exceeding the sensitivity of the compendial method. We present herein a NAT method for the detection of Mycoplasma that circumvents the need for live Mycoplasma spp. in the test procedure by instead being qualified using Mycoplasma spp. genomic DNA. We have demonstrated a lower limit of detection that exceeds the regulatory requirements set by Health Canada. This assay is now being used to screen clinical cell therapy products manufactured at our center.