MYCOPLASMA DETECTION BY PCR TEST

Mycoplasma can contaminate cell cultures and cost biotechnology companies lost time and reduce profits. The organism can severely reduce cell growth and metabolism, which will affect the field and possibly the activity of the recombinant protein. Mycoplasma is a unique group of microorganisms that fall in the category between the bacteria and viruses.

Currently, European and US regulations demand that CHO cells used to make recombinant therapeutic proteins are routinely tested for mycoplasma, a process that can take four to six weeks using conventional approaches such as broth/agar cultures and DNA staining of indicator cell cultures.

MQA developed a new method based on Polymerase Chain Reaction (PCR) that reduces the mycoplasma screening test to 1-2 days. MQA method is GMP validated using 6 different mycoplasma species. In extracts of mycoplasma DNA and in the presence of CHO cell with antibiotic, the assay limit of detection for six different mycoplasma species is 10 genomic copies. The assay is very specific to detect mycoplasma in the presence of other genomic DNA.
Cell line samples should be pre-cultured in the absence of mycoplasma active antibiotics for several days to maximize test sensitivity. Samples should be derived from cultures that are at 90-100% confluence (1.0 x 106 to 1.0 x 107cells). PCR inhibiting substances may accumulate in the medium of older cultures.