INDEL-Typing of <i>Yersinia pseudotuberculosis</i> Strains

Abstract

The aim of this study was to develop a new method of intraspecific genetic differentiation of Yersinia pseudotuberculosis, based on the detection of INDEL-markers using PCR. Materials and methods. Analyzed were 308 strains from the NCBI database and 15 strains sequenced within the frames of this study. The nucleotide sequences of the strains were determined using the MiSeq technology platform. The genomes of the strains sequenced in the work, as well as genomes from the NCBI database, were assessed using in silico PCR with 7 pairs of primers designed in the study. As a result of a comparison of genome-wide sequences of 22 Y. pseudotuberculosis strains from the NCBI database, using the author’s software (GenExpert), 7 INDEL-markers were selected that make it possible to effectively distinguish between strains of the causative agent of pseudotuberculosis. Based on these markers, 7 pairs of primers were designed and synthesized for the analysis of different strains using PCR. Analysis of 323 strains in PCR in silico and 70 strains in PCR in vitro allowed for dividing them into 30 genetic groups. Comparison of the results of PCR in silico and in vitro confirmed the possibility of using the proposed primers for intraspecific differentiation of Y. pseudotuberculosis. Based on the data obtained, a dendrogram reflecting the phylogenetic relations of different strains of Y. pseudotuberculosis was constructed. When analyzing the distribution of Y. pseudotuberculosis strains by various clusters and genetic groups, a number of patterns were revealed. Conducted in silico and in vitro PCR show that the proposed method of INDEL-typing can be used for intraspecific genetic differentiation of the causative agent of pseudotuberculosis.