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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| Authors: | Dmitriy V Volokhov Joseph George Sue X Liu Pranvera Ikonomi Christine Anderson Vladimir Chizhikov |
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| Institution: | (1) Center of Biologics Evaluation and Research, Office of Vaccines Research and Review, Division of Viral Products, Laboratory of Methods Development , US Food and Drug Administration , HFM-470, 1401 Rockville Pike , Rockville , MD , 20852 , USA;(2) American Type Culture Collection (ATCC) , 10801 University Boulevard , Manassas , VA , 20108 , USA |
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| Abstract: | Abstract
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. |
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