%0 Journal Article %T Differentiation and specific detection of Shigella genus using 16S Single Specific Primer PCR (16S SSP- PCR) %J Cellular and Molecular Research (Iranian Journal of Biology) %I Iraninan Biology Society %Z 2383-2738 %A Mollasalehi, Hamidreza %A Vahedipour, Narges %A Alimi, Mahrokh %D 2022 %\ 01/21/2022 %V 34 %N 4 %P 565-574 %! Differentiation and specific detection of Shigella genus using 16S Single Specific Primer PCR (16S SSP- PCR) %K 16S rRNA %K single specific primer PCR %K Bacterial diagnosis %K Shigella genus %R %X In this study, we developed a single specific primer PCR (SSP-PCR) method for specific identification of Shigella genus with the use of ribosomal RNA gene. Regarding high copy number and stability in cells, rRNA is one of the efficient targets to detect cells exclusively. 16S rRNA gene is considered as a standard biomarker for classifying and identifying prokaryotic cells. With respect to high sequence homology among 16S rRNA gene and shortness of specific heterogenic region in this gene, SSP-PCR is used to facilitate the specific identification of 16S rRNA gene in this study. In this method, only one specific primer is designed for amplification of target genes. 16S rRNA gene of different species of Shigella genus was aligned with 57 other related bacteria and then a single specific primer was designed in the V6 variable region. The amplicon was analyzed on a two-dimension agarose gel electrophoresis. The 263 bp band was considered as positive result in the tested samples and was observed for all species of Shigella (Sh. flexneri, Sh. boydii, Sh. dysenteriae, Sh. sonnei), whereas other species remained unidentified. The specificity of the assay was evaluated to be complete. Combination of targeting 16S rRNA and using SSP-PCR method, is an efficacious method for specific detection of bacteria in a minimum required time with a great ease. Among the applications are namely the early-phase diagnosis of bacterial infections in acute emergency conditions such as natural disasters and countries’ defense industry in the form of diagnostic kits for clinical laboratories. %U https://cell.ijbio.ir/article_1694_f00f199bd8c7c2ccb85bd8d6d72b4eb9.pdf