Genus- and species-specific identification of mycoplasmas by 16S rRNA amplification

F J van Kuppeveld, J T van der Logt, A F Angulo, M J van Zoest, W G Quint, H G Niesters, J M Galama, W J Melchers
1993 Applied and Environmental Microbiology  
Systematic computer alignment of mycoplasmal 16S rRNA sequences allowed the identification of variable regions with both genus-and species-specific sequences. Species-specific sequences of Mycoplasma collis were elucidated by asymmetric amplification and dideoxynucleotide sequencing of variable regions, using primers complementary to conserved regions of 16S rRNA. Primers selected for Mycoplasma pneumoniae, M. hominis9 M. fermentans, Ureaplasma urealyticum 9 M. pulmonis 9 M. arthritidis9 M.
more » ... arthritidis9 M. neurolyticum9 M. muris9 and M. collis proved to be species specific in the polymerase chain reaction. The genus-specific primers reacted with all mycoplasmal species investigated as well as with members of the genera Ureaplasma9 Spiroplasma9 and Acholeplasma. No cross-reaction was observed with members of the closely related genera Streptococcus9 Lactobacillus9 Bacillus9 and Clostridium or with any other microorganism tested. On the basis of the high copy number of rRNA, a highly sensitive polymerase chain reaction assay was developed in which the nucleic acid content equivalent to a single organism could be detected. The class Mollicutes consists of wall-less prokaryotes which are small in size and have unusually small genomes. More than 100 species have been isolated from vertebrates, plants, and insects. The largest group is formed by the genus Mycoplasma, of which more than 90 species have been described. The class Mollicutes contains five other genera
doi:10.1128/aem.59.2.655-.1993 fatcat:2p5v3cjq2nczjnjnnp4dkpemrm