How do you test for mycoplasma contamination?
The only way to confirm mycoplasma contamination is by routine testing using one or more special techniques, including direct growth on broth/agar, specific DNA staining, PCR, ELISA, RNA labeling and enzymatic procedures.
What is mycoplasma detection?
Mycoplasma testing includes a group of tests that either measure antibodies in the blood produced in response to a mycoplasma infection or detect the microbe directly through culturing or by detecting its genetic material (DNA) in a body sample. It is most often used to detect Mycoplasma pneumoniae (M.
How is cell culture contamination detected?
Bacterial contamination is easily detected by visual inspection of the culture within a few days of it becoming infected;
- Infected cultures usually appear cloudy (i.e., turbid), sometimes with a thin film on the surface.
- Sudden drops in the pH of the culture medium is also frequently encountered.
How common is mycoplasma contamination?
Mycoplasma contamination of cell cultures has been known for decades and disturbingly, has become widespread, threatening academic labs to biopharmaceutical production facilities. In fact, depending on the laboratory, anywhere from 10% to 85% of cell lines may be contaminated.
Where does mycoplasma come from cell culture?
There are three major sources leading to mycoplasma contamination of cell cultures in the laboratory: infected cells sent from another lab; contaminated cell culture medium reagents such as serum and trypsin; and laboratory personnel infected with M. orale or M. fermentans.
What is the best method to detect mycoplasma?
Mycoplasma contamination can be detected by polymerase chain reaction (PCR). PCR is easy, sensitive, specific, fast, reliable, efficient and costeffective. The PCR test is based on the detection of 16S rRNA molecules of the most common species of mycoplasma contaminating cell cultures.
Is mycoplasma inside a cell?
They are present in most of the cell culture facilities, tissue culture labs and every cell culturist has to deal with this problem. It is estimated that mycoplasma is responsible for up to 60% of the cell culture contamination (Uphoff, 2002). Mycoplasmas are considered to be one of the simplest and smallest bacteria.
How do you get rid of mycoplasma contamination?
Autoclaving the contaminated cell cultures is the best way to get rid of the infections. In the case of valuable cells contaminated by mycoplasmas, autoclave cannot be helpful and an elimination method should be used without harming the eukaryotic cells.
What is the source of contamination in my cell culture?
Culture media, bovine sera, reagents, and plasticware can also be major sources of biological contaminants. While commercial testing methods are much improved over those of earlier decades, it is paramount to use materials that are certified for cell culture use. Cross-contamination can occur when working with multiple cell lines at the same time.
What can be the cause for contamination of cell culture?
Researchers typically think of cell culture contamination in terms of microorganisms such as bacteria, fungi, viruses, and protozoa, but contamination can also be caused by chemical contamination as well as contamination by other cell lines – for example HeLa cells.
What is the genetic material in Mycoplasma?
The genetic material of mycoplasma consists of a naked circular chromosome of fibrilar DNA which is about 3 nm in thickness and 1000 x 10 6 daltons in weight (1 dalton = 1/16 the atomic weight of oxygen = 1.650 x l0 -24 g). This replicates in the same way as does the bacterial chromosome.
What is cell culture contamination?
Cell Culture Contamination. Cell culture contamination happens when the cells become infected with bacteria, mycoplasma, yeast and/or mold. Using aseptic technique is important in preventing this type of contamination.
