Mycoplasma contamination of cell lines is a common occurrence and may affect the cell line behaviour in a variety of ways. Contamination with mycoplasma is usually not obvious so cell lines should be frequently tested. Several commercially available kits for mycoplasma detection exist, however the Ph. Eur. culture method which can take several weeks to yield results is still considered to be the ‘gold standard’ method. There is therefore a need for rapid alternative methods with comparable sensitivity, specificity and species range. Here, we describe an internally-controlled Taqman-based real-time PCR assay for cell culture medium without the need for DNA extraction. The assay can detect less than 10 CFU of the most frequently encountered mycoplasma contaminants in mammalian cell cultures. The validated assay has the potential to be used as a routine test in the production of cell culture-based Biologicals.

细胞系的支原体污染是常见的现象，并可能以多种方式影响细胞系的行为。通常不明显感染支原体，因此应经常测试细胞系。存在几种用于支原体检测的可商购的试剂盒，但是Ph。Eur。可能需要数周才能产生结果的培养方法仍被认为是“金标准”方法。因此，需要具有可比的敏感性，特异性和种类范围的快速替代方法。在这里，我们描述了一种内部控制的基于Taqman的实时PCR分析方法，无需进行DNA提取即可用于细胞培养基。该检测方法可以检测不到10 CFU的哺乳动物细胞培养物中最常见的支原体污染物。