Investigation of fast methods for detection of mycoplasma contamination in cell lines in laboratory

Abstract

Objective To establish fast and reliable methods for detection of mycoplasma contamination in cell culture in laboratory. Methods Eleven cell lines were stained by DNA fluorescein stain, and the results of staining were analysed. Two pieces of highly conserved oligonucleotide sequences from mycoplasmal 16S rRNA of contaminating cells were selected and used as primers of mycoplasma. The mycoplasma contamination in cell culture was determined by PCR. A comparison of the results of determination was made between the two methods. Results Out of 11 cell lines, 2 ( 18.2% ) gave a positive result with DNA fluorescein staining method, and 4 (36.4%) gave a positive result with polymerase chain reaction (PCR) method. A comparison between the two methods indicated that those cell lines positive by DNA fluorescein staining were also positive by PCR. Conclusions The above-mentioned two detection methods can effectively detect mycoplasma contamination in cell lines. The sensitivity of DNA fluorescein staining is lower than that of PCR, but the cost of PCR is higher. Combined use of the two detection methods, for example, the cell lines being suspectedly positive by DNA fluorescein staining are detected by PCR, can raise positive detection rate, and lower cost. Key words: Mycoplasma; Polymerase chain reaction; DNA fluorescein staining; Contamination