RAPID IDENTIFICATION OF SACCHAROMONOSPORA STRAINS BY MULTIPLEX PCR USING SPECIES-SPECIFIC PRIMERS WITHIN THE 16S RIBOSOMAL-RNA GENE

A multiplex PCR technique for the rapid identification of Saccharomono spora strains was developed. Primers for the four validly described Sa ccharomonospora species were designed by aligning previously published 16S rRNA sequences. These primers were found to be species-specific t hrough the application of a specificity test. Four species-specific pr imers were added to a single reaction tube together with a universal r everse primer corresponding to the 3' terminus of 16S rRNA genes. The representative strains of the four species could be differentiated by the PCR products characteristic of each species. The five strains of ' 'Saccharomonospora caesia'' yielded an identical PCR profile to that o f Saccharomonospora azurea K161(T) as supposed from the same 16S rRNA sequence between S. azurea K161(T) and the strains of ''S. caesia''. T he five strains of Saccharomonospora glauca and the five strains of Sa ccharomonospora viridis showed identical results to those of correspon ding representative strains. It is now possible to rapidly identify th e strains of the genus Saccharomonospora using this multiplex PCR assa y. This method was found to be simple, reproducible and species-specif ic.