As mollicutes usually cause diseases of the respiratory or the urogenital tract, where the spectrum of pathogens to be considered is large, considerable interest has been focused on the simultaneous detection of several molli-cutes, or mollicutes and other pathogens, by the same assay. Although the regions V2 and V3 of the 16S rDNA display sufficient interspecies differences for species-specific detection, conserved sequences of the 16S rDNA gene enable detection of most Mycoplasma spp., Urea-plasma spp., Spiroplasma spp., and Acholeplasma spp. by a single assay using the primer pair GPO-1/MGSO. All mollicutes, which have been isolated from humans, are listed in Table 1.
A recently described seminested PCR based on this assay using the additional primer My-ins provides improved sensitivity. This assay has been shown to allow amplification of M. genitalium, M. hominis, Mycoplasma faucium, U. urealyticum, and U. parvum specific sequences.
A variety of assays for simultaneous detection of mollicutes and other pathogens have been described, e.g., a multiplex real-time PCR assay to detect M. pneumoniae, Chlamydia pneumoniae, and Legionella pneumophila in respiratory samples. In addition, a multiplex RT-PCR for the detection of M. pneumoniae and eight additional respiratory pathogens has been constructed.1-18-1 Simultaneous detection of M. hominis together with the bacterial vaginosis-associated pathogens Lactobacillus spp. and Gardnerella vaginalis by real-time PCR has been described.
When cell cultures are to be tested for mycoplasma contamination, a convenient alternative to PCR (using GPO-1/MGSO) is a commercially available ELISA kit for antigen detection.
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