聚合酶链式反应快速检测在细菌类微生物污染细胞培养液中的应用价值

The Application Value of PCR Rapid Detection in Cell Culture Medium Contaminated by Bacteria and Microorganisms

目的分析聚合酶链式反应(PCR)快速检测在细菌类微生物污染细胞培养液中的应用价值。方法本研究材料选择人为污染海拉细胞系(HeLa细胞),细胞类微生物通用引物使用16 S核糖体RNA(16 S rRNA)基因序列,对16 S rRNA基因序列片段进行PCR扩增,设定PCR检测培养细胞污染方式,并用该方法对本次细胞株的污染状况进行详细检测。结果在人为污染大肠杆菌、绿脓杆菌、白色葡萄球菌的HeLa细胞的培养上清中,均存在扩增大小片段,并且与目的片段相符合。在本研究中的20个细胞株培养上清里,扩增出大小片段的有5株。结论本研究建立16 S rRNA基因序列通用引物PCR法,能够快速有效检测细胞培养中细菌类微生物污染,对早期发现污染有重要的应用价值。

Objective To analyze the application value of rapid PCR detection in cell culture medium contaminated by bacteria and microorganisms.Methods Materials for this study a human contaminant HeLa cell line(Hela cells)was selected,and the cell class microbial universal primers used 16 S ribosomal RNA(16S rRNA)gene sequences,PCR amplification of 16 S rRNA gene sequence fragments,set PCR mode to detect the contamination of cultured cells,and using this method,the contamination status of this cell line was examined in detail.Results In the culture supernatant of HeLa cells artificially contaminated with Escherichia coli,Pseudomonas aeruginosa,Staphylococcus albicans,both amplified size fragments were present and corresponded to the fragment of interest.Of the 20 cell strain culture supernatants in this study,five amplified large and small fragments.Conclusion This study established a universal primer PCR method for 16S rRNA gene sequence,which can quickly and effectively detect bacterial microbial contamination in cell culture,and has important application value for early detection of contamination.