Mycoplasma detection may not be the most rewarding task, but the latest products are ensuring that you’re not wasting your time. These fresh approaches and designs help you sleep easy at night, knowing that you’ve uncovered any and all of these bacterial perpetrators. As with domestic spying, the newest kits and reagents strive to prevent sabotage of your cell culture’s otherwise perfect world.
In humans, species of the Mycoplasma phylum are involved with a host of diseases, including pneumonia and other upper respiratory infections, chronic fatigue syndrome, Gulf War syndrome, and diseases of the blood, joints, central nervous system, liver, pancreas, and cardiovascular system. It’s no wonder that these tiny prokaryotes can wreak havoc in your cell cultures by altering cell growth and metabolism, nucleic acid synthesis, membrane antigenicity, enzyme kinetics, and other vital functions and processes. Unfortunately, up to 87% of all cell cultures are contaminated with Mycoplasma, which are so small that they often pass through the filters routinely used for sterilizing culture reagents. And because the bacteria don’t have cell walls, antibiotics have little effect.
Chances are that sterile techniques won’t preclude you from having to perform that periodic Mycoplasma check. The many routes of infection—bench surfaces, dust, skin, hair, clothing, laminar flow hoods, and incubators—can overwhelm the most careful scientist. Even more frustrating is that Mycoplasma detection is not readily apparent. The culture doesn’t become cloudy. The pH doesn’t change. All the while, you could be unwittingly collecting erroneous data.
Today’s Mycoplasma detection products leave you with little excuse to forgo periodic checks. It’s no longer a day- or week-long process involving tedious DNA staining and microbiological cultures. Mycoplasma detection can now be performed in a shorter time than the average morning commute. Within 15 minutes, some kits can tell you if your cultures are contaminated and by which bacterial species. Kits include those employing PCR, biochemical assays, and immunofluorescence and ELISA-type immunoassays.
Primers are the special ingredients in kits that use PCR detection methods. These primers usually target the 16S and/or 23S highly conserved ribosomal RNA genes. Most kits rely on primers designed to detect several species but safeguarded against specificity for Escherichia coli DNA, which often contaminates recombinant Taq DNA polymerase. Gels reveal any bands indicating the presence of Mycoplasma, even in the tiniest amounts.
The biochemical assay kits exploit enzymes unique to Mycoplasma. Introduction of the respective substrates and measuring any enzyme action can point to contamination. Immunofluorescent kits provide Mycoplasma-specific antibodies that carry a probe for fluorescent or colorimetric detection. Strepavidin-conjugated antibodies give you the ability to amplify the signal, which helps to root out contamination that might otherwise remain hidden. Or, you might prefer microarrays that feature Mycoplasma-specific oligonucleotides.
Most every kit, such as those below, is practically guaranteed to have you breathing a sigh of relief. These products represent the great selection you’ll find on the market, outside of wiretapping. Best of all, monitoring for Mycoplasma is completely warranted.
