Open AccessDetection of bacterial and mycoplasma contamination in cell cultures by polymerase chain reaction
Author(s) -
Spaepen Marijke,
Angulo Alexander Fidèle,
Marynen Peter,
Cassiman JeanJacques
Publication year - 1992
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1992.tb05547.x
Subject(s) - polymerase chain reaction , biology , mycoplasma , primer (cosmetics) , microbiology and biotechnology , 16s ribosomal rna , bacteria , mycoplasmataceae , mycoplasma hominis , mollicutes , dna , gene , virology , genetics , chemistry , organic chemistry
A fast and simple method to detect bacterial and especially mycoplasma contamination in tissue culture by means of polymerase chain reaction (PCR) amplification is described. In a first step the universal primer pairs P1/P2 (190‐bp fragment) and P3/P4 (120‐bp fragment) directed to different conserved parts of the prokaryotic 16S rRNA gene are used. A positive signal after amplification on cell culture DNA with these primers provides an indication of bacterial infection. Using the internal primers IP1, IP3 and IP′3 complementary to a part of the V4 and V8 variable regions of the 16S rRNA gene, in combination with a universal primer, cultures contaminated with mycoplasma could be identified. Six mycoplasma species, typical contaminants in tissue cultures, were investigated: Mycoplasma orale, M. fermentans, M. arginini, M. hyorhinis, M. hominis and Aeromonas laidlawii . This mycoplasma test is an easy, specific and sensitive assay which should be extremely useful in any tissue culture setting.