Mycoplasma has garnered interest due to its ability to cause contamination of cell lines used in research and manufacturing bioproducts. Without a cell wall, mycoplasmas are invisible to the naked eye. In the past, they were commonly used as sources of animal sera. Mycoplasmas can be found in the pipetting through the mouth. They are highly resistant to antibiotics and can only be detected by the presence of cell membranes and supernatants.
Although the frequency and impact of their contamination have been discussed, the incidence of single mycoplasma infections is still high, ranging from 15 to 35% globally. Mycoplasma contamination can be challenging to detect in a lab. It can affect up to 30% of the global cell lines used in research.
Methods of Identifying Mycoplasma Contamination
The bacterial culture method is one of the most widely used Mycoplasma Detection in cell culture. This test involves the addition of a sample of cell culture supernatant to a liquid medium.
The positive results of this test will show colonies that are similar to fried eggs with a diameter of 100 to 400 m. This method is susceptible to detecting the presence of mycoplasma contamination.
Although some strains of mycoplasma can’t be cultivated in this method, a certain number can still thrive in cultivation media.
The European Pharmacopeia has approved a second Mycoplasma Detection method involving staining a sample’s DNA with fluorophores. This method is commonly used for rapid and easy tests.
If the cell culture conditions are not good, the results of the direct DNA staining method may be misinterpreted. This is why the use of indicator cell lines is helpful.
The polymerase chain reaction is a fast and sensitive Mycoplasma Detection method to identify mycoplasma contamination. It uses 16S rRNA molecules to identify the most common mycoplasma species.
The specificity of primers used in this method should be narrow enough to prevent the amplification of other bacteria.