How do you identify mycoplasma contamination in cell culture?

How do you identify mycoplasma contamination in cell culture?

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.

What does mycoplasma do to cell culture?

Mycoplasma compromises cell culture-based experiments in the following ways: Competes for nutrients – hinders cell growth and proliferation. Exposes cells to unwanted metabolites. Alters levels of protein, RNA, or DNA synthesis.

Can you see mycoplasma under light microscope?

Mycoplasma cells are very small bacteria therefore they cannot be detected by visual inspection using a visible light microscope and, thus, can remain unnoticed in the cell cultures for long periods.

How do you identify contamination in cell culture?

Bacterial contamination is easily detected by visual inspection of the culture within a few days of it becoming infected;

  1. Infected cultures usually appear cloudy (i.e., turbid), sometimes with a thin film on the surface.
  2. Sudden drops in the pH of the culture medium is also frequently encountered.

How is mycoplasma treated in cell culture?

There are three classes of antibiotics that kill mycoplasma when used at relatively low concentrations: tetracyclines, macrolides and quinolones. Tetracyclines and macrolides block protein synthesis by interfering with ribosome translation, whereas quinolones inhibit replication of mycoplasma DNA.

What is mycoplasma test?

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.

What is contaminated culture?

A culture in which bacteria from a foreign source have infiltrated the growth medium. See also: culture.

Can you see mycoplasma on Gram stain?

Mycoplasma pneumoniae is the pathogen most commonly associated with disease in humans. It is a short rod with no cell wall; therefore, it is not visible on the Gram stain. It can be isolated on media supplemented with serum.

Can mycoplasma spread in incubator?

The mycoplasmas enter the cell culture through various sources that are difficult to trace. These include the laboratory personnel, the serum, the cell culture media, water baths, incubators, etc. Mycoplasmas can spread from these sources through cross-contamination and due to poor lab techniques.

What is contaminated culture in microbiology?

What is the best antibiotic for mycoplasma?

Medication Summary The second-generation tetracyclines (doxycycline) and macrolides are the drugs of choice. Macrolide resistance has been reported in several areas of the world, but most experts agree that macrolides are the antibiotics of choice for treating M pneumoniae infections in adults and children.

How is mycoplasma diagnosed?

Standard PCR is currently the method of choice for direct detection of M. pneumoniae. PCR has replaced hybridization and direct antigen detection because of its higher sensitivity. During recent years, several PCR-related methods have been applied for research purposes.

Is it possible to detect Mycoplasma contamination in cell cultures?

Unfortunately, it is challenging to quickly and reliably detect mycoplasma contamination in cell cultures. One reason is that mycoplasma contamination does not typically trigger the turbidity changes that are common with other types of bacterial or fungal contamination.

What are the signs and symptoms of Chronic Mycoplasma infection?

Chronic mycoplasma infections might manifest themselves with decreased rate of cell proliferation, reduced saturation density, and agglutination in suspension cultures; however, the only assured way of detecting mycoplasma contamination is by testing the cultures periodically using fluorescent staining (e.g.,…

Does Mycoplasma contamination slow down cell metabolism?

Even though the mycoplasma contamination does not slow down cell metabolism, it may contaminate the final product (such as a vaccine) resulting in the loss of the batch ( 26 ). Mycoplasmal infection of cell cultures might often linger for an extended period of time without noticeable cell damage ( 7, 23, 27 ).

How does mycofluor™ stain Mycoplasma?

In live cells, the MycoFluor™ reagent does not have access to the nuclei, but readily stains the mycoplasma associated with the outside of cells (panel C). The images were obtained using 365 nm excitation and a 100/1.3 Plan Neoflaur® (Zeiss) objective lens coupled with a 450 ± 30 nm bandpass filter.