Mycoplasmas are small, round or filamentous prokaryotic organisms that lack cell walls. It has been estimated that at least 15%-35% cell cultures are contaminated with mycoplasma. Mycoplasma contamination can modify many aspects of cell physiology without causing obvious medium cloudiness and apparent effect on cell growth. Mycoplasma contamination affects up to 80% of continuous cell cultures. If undetected, mycoplasma contamination can have significant effects on the quality and reliability of your cell culture preparations. Due to their small size and deformability, mycoplasmas can pass through 0.22µm filter. The lack of cell wall makes mycoplasma unresponsive to common antibiotics that target cell wall synthesis. It is essential that all cell stocks and new cultures entering a facility are tested for the presence of mycoplasma and all cell cultures should be tested routinely (e.g. once every 2-3 months) in order to maintain a mycoplasma-free environment.

Many methods are available for detection of mycoplasma, including isolation in broth/agar culture, direct or indirect fluorescence staining, ELISA, immunostaining, direct or indirect PCR. Among those methods, direct PCR is the most sensitive, specific and convenient method when the primer design is optimized.

The MycoDect™ Mycoplasma Detection Kit is designed to detect mycoplasma infection in cell cultures in less than two hours. It can detect mycoplasma from both cell lysates and cell culture media. The sensitivity is up to 10-20 copies of target DNA, which translates to less than 10 mycoplasma species. This kit detects the top five common species of mycoplasma, M. Hyorhinis, M. Arginini, M. Orale, M. Fermentans, A. laidlawii, which represent 98% of tissue culture infections. It can also detect Mycoplasma, Acholeplasma, Ureaplasma and Spiroplasma from other genera.