基于 PCR 的检测和消除各种哺乳动物细胞系中的支原体感染：本地经验。
PCR-based detection and eradication of mycoplasmal infections from various mammalian cell lines: a local experience.
机构信息
National Cell Bank of Iran, Pasteur Institute of Iran, Tehran, Iran.
出版信息
Cytotechnology. 2009 Dec;61(3):117-24. doi: 10.1007/s10616-010-9252-6. Epub 2010 Feb 6.
A total of 200 cell lines including different human, monkey, mice, hamster and rat cell types were examined for mycoplasma infection status. PCR assay using generic-specific universal primers showed that 40 (20%) of the cell lines are contaminated with mycoplasma. Employment of species-specific primers within these infected cell lines revealed infection with M. hyorhinis (42.5%), M. fermentas (37.5%), M. arginini (37.5%), M. orale (12.5%) and A. laidlawii (7.5%). A number of the cultures were coinfected with 2 or 3 different species. Contaminated samples were treated with BM-Cyclin, Ciprofloxacin and mycoplasma removal agent (MRA). Mycoplasma eradication was subsequently checked by PCR following 2 weeks continuous culture of treated cells in antibiotic free culture medium. Mycoplasmal infections were eradicated in 100, 70 and 42% of infected cell lines when the samples were treated with BM-Cyclin, MRA and Ciprofloxacin, respectively. However, 12% (BM-Cyclin), 62.5% (MRA) and 82.5% (Ciprofloxacin) of mycoplasma regrowth was observed 4 months after the treatment. Notably, the risk of spontaneous culture death was 17.5, 12.5 and 0% for BM-Cyclin, MRA and Ciprofloxacin, respectively.
共检测了 200 种细胞系，包括不同的人、猴、鼠、仓鼠和大鼠细胞类型，以确定支原体感染状况。使用通用特异性引物的 PCR 检测显示，40 种（20%）细胞系受到支原体污染。在这些受感染的细胞系中使用种特异性引物，发现感染了 M. hyorhinis（42.5%）、M. fermentas（37.5%）、M. arginini（37.5%）、M. orale（12.5%）和 A. laidlawii（7.5%）。一些培养物同时感染了 2 种或 3 种不同的物种。用 BM-Cyclin、环丙沙星和支原体去除剂（MRA）处理污染的样品。随后在无抗生素的培养基中连续培养处理后的细胞 2 周，通过 PCR 检查支原体的消除情况。用 BM-Cyclin、MRA 和环丙沙星处理感染细胞系时，支原体的消除率分别为 100%、70%和 42%。然而，处理后 4 个月观察到支原体再生长的比例分别为 12%（BM-Cyclin）、62.5%（MRA）和 82.5%（环丙沙星）。值得注意的是，BM-Cyclin、MRA 和环丙沙星处理后自发培养死亡的风险分别为 17.5%、12.5%和 0%。
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