Identification of clinical Proteus isolates by using rpoB gene sequence analysis

Abstract

: Some bacterial genera and species could not be identified by 16S rRNA sequencing, so the rpoB gene has been suggested to be the alternative because the sequence of the rpoB gene is more distinguished than the 16S rRNA gene to discriminate various species of bacteria. Thus, this study has been designed to show the usefulness of the rpoB gene in accurately identifying Proteus clinical isolates. Fifty bacterial isolates were collected and identified phenotypically according to the culture, microscopic examination and biochemical tests. VITEK 2 compact system was used to confirm identification. Genotypic identification was performed after DNA extraction for 10 selected isolates and amplification with rpoB gene-specific primer and gel electrophoresis. The products were detected with a (1090 bp) molecular size band, which was sent for Sanger sequencing using an ABI3730XL automated DNA sequencer, and data were analyzed and compared with standard sequences in GenBank. It has been concluded that identifying and discriminating Proteus species by rpoB gene sequence analysis is more correct and accurate. Keywords: 16S rRNA, rpoB gene, Proteus spp. identification, sequencing