关键词: 中国仓鼠卵巢细胞, 支原体, PCR反应

Abstract: For the detection of mycoplasma contamination in Chinese hamster ovary cells (CHO) cell culture, a rapid, accurate, and stable PCR detection method was established. A housekeeping gene encoded glyceraldehyde 3-phosphate dehydrogenase was used as a reference gene, and the internal reference primers and primers for mycoplasma detection were placed in a PCR system simultaneously. The concentration ratio of two kinds of primers was optimized as 5 μmol/L reference gene primers and 10 μmol/L mycoplasma detection primers. PCR method had consistent results with DNA staining method, with less false negative result and short detction time, showing that it could meet the demand for mycoplasma contamination detection in the production process.

Key words: Chinese hamster ovary cell, mycoplasma, PCR reaction