The PCR primer sets used in McCT are directed to the highly conserved 16S rRNA coding region in the Mycoplasma genome. This allows for amplification of all Mycoplasma and Acholeplasma species usually encountered as contaminants in cell cultures. Moreover, the McCT assay determines the genotype of Mycoplasma and Acholeplasma species by specific hybridisation, such as M. arginini, M. fermentans, M. orale, M. hyorhinis, M. hominis, M. salivarium, M. synoviae, M. pirum, M. gallisepticum, M. pneumoniae, M. yeatsii, and Acholeplasma laidlawii.
Mycoplasma specification enables the identification of contamination sources:
The Central Commission for Biological Safety (ZKBS) at the Robert-Koch Institute (RKI) has assessed the risk for cell lines which were immortalised with Epstein-Barr Virus (EBV) also known as Human Herpes Virus 4 (HHV4). Cell lines that are immortalised due to an EBV infection must be used in S2. The cell lines MEC-1/MEC-2, EHEB, Granta 519, JVM-2/JVM-3 and LCL-WEI were assigned to S2. EBV positive cell lines are allowed in S1 only if a sequence defect of a gene essential for lytic replication is detected.
Multiplexion´s Multiplex Cell Contamination Test tests for the EBV DNA polymerase processivity factor in ORF59.
For further information:
The Central Commission for Biological Safety (ZKBS) at the Robert-Koch Institute (RKI) has recommended testing of cell lines for the Squirrel Monkey Retrovirus (SMRV) that is able to infect a broad range of cell lines. Like all retroviruses, SMRV poses a risk for insertional mutagenesis. In addition, SMRV contamination may lead to a complementation of recombinant, replication-deficient retroviruses in cell lines. Contaminated cells should either be inactivated or used in S2 only. The ZKBS further asks to get notified if a cell line is tested SMRV positive.
For further SMRV information