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Updated: Jun 16, 2026
Optimized PCR-based Detection of Mycoplasma
Published on: June 20, 2011
Cross comparison of rapid mycoplasma detection platforms
Bill Lawrence1, Houman Bashiri, Houman Dehghani
1Biosafety Development Laboratory, Cellular Resources, Amgen, 1201 Amgen Court West, Seattle, WA 98119, USA. bill.lawrence@amgen.com
Abstract:
The use of animal and plant derived raw materials in mammalian cell culture processes may provide a possible route of entry for adventitious contaminants such as mycoplasma. Mycoplasma contaminations of cell culture represent a serious challenge to the production of biotechnology derived therapeutics. The slow growing nature of mycoplasma can disguise their infection of cultures since cells may continue to proliferate, though at reduced levels and with lesser output of engineered protein. Rapid identification of mycoplasma contaminated cell cultures and materials enables a faster response time to prevent the spread of the contamination. We describe here the comparison of different mycoplasma detection methods: two nucleic acid-based technologies, the standard mycoplasma culture procedure, and a hybrid culture-quantitative PCR assay. In this study, a cell line infected with two species of mycoplasma was used to compare the different detection methods. Our data demonstrates that the two nucleic acid-based techniques are robust methods for detection of mycoplasma and have similar detection capability. In contrast, no mycoplasma was detected in the standard culture assay or in a hybrid culture-quantitative PCR assay. This shows a potential limitation of the culture assay that relies on the ability of mycoplasma to grow in broth media.
Insights
Rapid detection of mycoplasma contamination in cell cultures is crucial for biopharmaceutical production. Nucleic acid-based methods proved superior to culture-based assays for identifying mycoplasma infections.
Area of Science:
- Biotechnology
- Cell Biology
- Microbiology
Background:
- Animal and plant-derived raw materials in mammalian cell culture can introduce adventitious contaminants like mycoplasma.
- Mycoplasma contamination poses a significant threat to the production of biotherapeutics, often remaining undetected due to slow growth.
- Early detection of mycoplasma is vital to prevent widespread contamination and ensure product safety.
Purpose of the Study:
- To compare the efficacy of different mycoplasma detection methods in mammalian cell cultures.
- To evaluate nucleic acid-based technologies against standard culture procedures and a hybrid assay.
Main Methods:
- Comparison of two nucleic acid-based detection techniques.
- Assessment of a standard mycoplasma culture procedure.
- Evaluation of a hybrid culture-quantitative PCR assay.
- Utilized a cell line intentionally infected with two species of mycoplasma.
Main Results:
- Nucleic acid-based techniques demonstrated robust and comparable detection capabilities for mycoplasma.
- The standard culture assay and the hybrid culture-quantitative PCR assay failed to detect mycoplasma in the infected cell line.
- Highlighted a limitation in culture-based methods dependent on mycoplasma's ability to grow in broth media.
Conclusions:
- Nucleic acid-based methods are highly effective for rapid and reliable mycoplasma detection in cell cultures.
- Standard culture methods may not be sensitive enough for detecting certain mycoplasma infections, potentially leading to false negatives.
- The findings underscore the importance of employing advanced detection technologies to safeguard biopharmaceutical manufacturing processes.
