Mycoplasma Detection
Introduction
Mycoplasma contamination of cell culture systems continues to present major problems for basic research as well as for the manufacturing of bioproducts. It has been shown that mycoplasmas produce a variety of effects on cultured cells (e.g. changes in metabolism, immunologic or biochemical properties, growth, viability, etc.). Since mycoplasma infection in cell cultures is normally a chronic infection which may not be obvious by visual inspection or light microscopy, routine periodic screenings with a high sensitive method is an important evaluation of cell cultures. A variety of tests to detect mycoplasmas in cell cultures have been developed (e.g. fluorochrome staining of DNA, monitoring toxic metabolites, culture method, etc.) but each method shows certain disadvantages. The major drawback of those methods is non-specificity or a very time-consuming procedure.
Methodology employed
The methodology used to detect possible contamination with Mycoplasma is PCR-RT (Real Time PCR). This reaction is a process of PCR amplification and simultaneous detection of DNA amplified in "real time", such detection may be performed using different fluorescent elements, such as probes or intercalating agents that evidence the amount of DNA present in each amplification cycle.
The advantages of using Real Time PCR are:
- Homogeneous system that avoids manipulation of amplified DNA, escaping the serious problem of contamination with it.
- Speed of results.
- Wide dynamic range of detection.
- High sensibility.
- Minimum variation in the results, giving a reliable and accurate detection.
Detection limit
10 copies of DNA target / reaction
Delivery of results
7 working days
An additional service that Cromátida offers is the identification of Mycoplasma strain with which your sample is contaminated.
This is not only beneficial at the time of choosing a specific treatment for the removal of the contaminant strain, but also enables the evaluation of the possible source of contamination, e.g. Human Mycoplasma from operators (M.fermentans) , Mycoplasma from bovine serum (M.arginini), etc ...
As a global advantage of great importance it is worth noting that all of the above services enable the evaluation and generation of a preventive strategy and a work control in different areas that will improve the quality of work and avoid future contaminations.
Delivery of results
14 working days