Sequencing of the intergenic 16S-23S rRNA spacer (ITS) region of Mollicutes species and their identification using microarray-based assay and DNA sequencing
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We have completed sequencing the 16S-23S rRNA intergenic transcribed spacer (ITS) region of most known Mycoplasma , Acholeplasma , Ureaplasma , Mesoplasma , and Spiroplasma species. Analysis of the sequence data revealed a significant interspecies variability and low intraspecies polymorphism of the ITS region among Mollicutes . This finding enabled the application of a combined polymerase chain reaction–microarray technology for identifying Mollicutes species. The microarray included individual species-specific oligonucleotide probes for characterizing human Mollicutes species and other species known to be common cell line contaminants. Evaluation of the microarray was conducted using multiple, previously characterized, Mollicutes species. The microarray analysis of the samples used demonstrated a highly specific assay, which is capable of rapid and accurate discrimination among Mollicutes species.
We would like to express our appreciation to Dr. Maureen K. Davidson of the Mollicutes Collection at Purdue University, West Lafayette, IN, USA, for kindly providing us with Mollicutes species; Dr. J. Lindsay Oaks of the Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA, USA for kindly providing us with genomic DNA of Mycoplasma vulturii strain Gb-V33; and Dr. Michael Klutch for his assistance in sequencing.
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