Reliable, PCR-based mycoplasma detection
Douglas McKenzie Heinz Miller Ian Colbert
Stratagene introduces the Mycoplasma PCR Primer Set Plus, a kit that uses a PCR-based method which specifically amplifies an 874-bp PCR product from a single copy gene. In addition, the kit contains an internal control template that serves as a positive control for PCR. The kit offers a rapid and sensitive means to determine information about mycoplasma (and related acholeplasma) species. The kit expands on Stratagenes commitment to provide reagents that detect mycoplasma infection in cell cultures.
Mycoplasmas (and acholeplasmas) are wall-less eubacteria that exist as parasites within eukaryotic cells. As the smallest, simplest, and most primitive forms of self-replicating prokaryotes, they consist of only three major componentsa plasma membrane, ribosomes, and a double-stranded genome of approximately 580 kb. Because of their limited synthetic capacity, these organisms depend on host cells as a source of nutrients.
Mycoplasmas pose a recurring problem for investigators who use cells grown in culture. When mycoplasmas infect cell cultures, they dramatically effect virtually every aspect of cellular metabolism, without any apparent influence on cellular morphology. Often, only an increased cell doubling time indicates culture contamination. To avoid these artifactual effects, cell lines should be periodically evaluated.
The first protocols developed to detect mycoplasma infection in cell lines
required unwieldy probe technologies and time-consuming cell culture methods
using indicator cells and suicide substrates.1 Subsequently,
Stratagene introduced the first PCR-based method for mycoplasma detection.2
While other companies have since int