Isolation and Identification of Pathogenic Yersinia Enterocolotica Using 16 SrRNA from Children in AboRish Hospital, Egypt

Abstract

Foodborne diseases are a widespread and growing public health concern in developed and developing countries. Isolation of Yersinia enterocolitica is include enrichment followed by streaking on selective agar such as cefsulodin, irgasan and novobiocin medium (CIN) Antimicrobial susceptibility determination of Yersinia enterocolitica strains was performed according to the NCCLS Standards. The use of PCR has shown to be a sufficient standard of specific identification and is not time consuming. DNA Sequencing of 16S rRNA gene was conducted in both directions and a consensus sequence of 1476 bp was used for nucleotide (nt.) analysis. The original sequences were trimmed to remove ambiguous nt. sequences usually exist in the beginning of the sequencing reaction. Partial DNA sequences were submitted to GenBank database and obtained accession numbers; MK168055 and MK168056 for Y.Ent-1/EGY018 and Y.Ent-2/EGY018 strains, respectively. Identification of homologies between nucleotide sequence of the studied Yersinia Spp. isolates and others published in GenBank was done using BLAST 2.2 search program (National Center for Biotechnology Information “NCBI” http://www.ncbi.nlm.nih.gov/). Both Y.Ent-1-EGY2018 and Y.Ent-2-EGY2018 strains revealed high similarity (96.6%) between each other. Sequence analysis of Y.Ent-1- EGY2018 and Y.Ent-2-EGY2018 showed 96.3-97.6% and 97.1-98.5% identity (respectively) when compared to other available Yersinia enterocolitica partial sequences of 16S rRNA gene published in GenBank. Phylogenic analysis of 16S rRNA gene partial sequence showed that Y.Ent-2-EGY2018 strain was isolating in same cluster of Yersinia enterocolitica but exists in different branches. However, Y.Ent-2EGY2018 strain is present alone in a separate cluster.