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Updated: Feb 3, 2026
06:01
Optimized PCR-based Detection of Mycoplasma
Published on: June 20, 2011
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Evaluation of mycoplasma removal reagents using qPCR-based quantification
Yueyang Lai1, Xuebo Xu1, Ruiying Yan1
1School of Life Sciences and the State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing, China.
Analytical Biochemistry
|October 19, 2018
Summary
Effective mycoplasma removal reagents Plasmocure and Plasmocin work within a week. Sequential treatment with Plasmocure and Primocin prevents mycoplasma recontamination, with qPCR methods quantifying contaminants per cell.
Area of Science:
- Cell Biology
- Microbiology
Background:
- Mycoplasma contamination is a common issue in cell cultures, potentially affecting experimental results.
- Reliable methods for detection and eradication are crucial for maintaining cell line integrity.
Purpose of the Study:
- To evaluate the efficacy of different mycoplasma removal reagents.
- To establish a quantitative method for assessing mycoplasma load.
- To determine optimal treatment strategies for mycoplasma eradication and prevention.
Main Methods:
- Evaluation of mycoplasma removal reagents using nuclear staining, DNA gel electrophoresis, and quantitative PCR (qPCR).
- Development of qPCR-based methods to estimate mycoplasma load per cell.
- Assessment of reagent efficacy, resistance development, and recontamination after treatment withdrawal.
Main Results:
- Plasmocure and Plasmocin demonstrated efficacy within one week.
- Prolonged Plasmocin treatment resulted in mycoplasma resistance.
- Reagent withdrawal led to mycoplasma reoccurrence within two days.
- Primocin effectively prevented recontamination.
- qPCR methods quantified 300-400 mycoplasma contaminants per cell in untreated contaminated cells.
Conclusions:
- Sequential treatment with Plasmocure followed by Primocin is recommended for effective mycoplasma control.
- Continuous use of a prevention reagent is imperative to avoid recontamination.
- Quantitative PCR provides a valuable tool for assessing mycoplasma burden and treatment effectiveness.
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