摘要
目的 检测细胞培养中细菌类(包括支原体)微生物的污染情况。方法 用细胞培养中常见的培养物人为污染HeLa细胞,设计细菌类微生物16SrRNA基因序列通用引物,PCR扩增16SrRNA基因序列片断,建立PCR检测培养细胞污染的方法,并用该方法检测本室保存的细胞株的污染情况。结果 人为污染绿脓杆菌、大肠杆菌、白色葡萄球菌和支原体M .fermentans的Hela细胞的培养上清中均扩增出大小的片段,与目的片段相符。本室所收藏的15个细胞株有3株的培养上清中扩增出大小的片段。结论 本实验建立了16SrRNA基因序列通用引物PCR法,可用于快速检测培养细胞中细菌类微生物的污染。
Objective To investigate a fast and simple method to detect bacterial contaminations including Mycoplasma contamination in cell cultures. Methods HeLa cells were used for propagation P. aeruginosa, E. coli, S. epidermidis and M. fermentans, respectively. The suitability of PCR based on the universal primers for the 16S rRNA gene sequences of prokaryotic cells for the detection of bacterial contamination in HeLa cell cultures was investigated. Then this method was used to test 15 cell strains in our laboratory. Results Nearly 1 500 bp region was amplified from all of the HeLa cell cultures contaminated by P. aeruginosa, E. coli, S. epidermidis or M. fermentans. Of the 15 cell strains in our laboratory, 3 were tested positive using this method. Conclusion The PCR method based on the universal primers for the 16S rRNA region of prokaryotic cells might be used for the rapid detection of bacteria in cell cultures.
出处
《局解手术学杂志》
2005年第2期88-90,共3页
Journal of Regional Anatomy and Operative Surgery
基金
国家自然科学基金资助项目 (3 0 3 0 0 3 0 3 )
第三军医大学创新研究基金
关键词
细胞培养
细菌
污染
PCR
cell culture
bacteria
contamination
PCR
