The use of the HRM method for identifying possible mutations in the ymoA gene region and evaluating their influence on the enterotoxic properties of Y. enterocoliticastrains

Abstract

BackgroundThe yst gene that encodes the production of Yst enterotoxins is one of the most important and reliable virulence markers. Its ability to produce Yst has been demonstrated in pathogenic strains isolated from clinical cases of yersiniosis with diarrhea. However, not all yst positive strains produce enterotoxins. According to some authors, Yst production can be restored in a silent strain by ymoA mutation. In this study, the HRM method was applied to identify ymoA single nucleotide polymorphism with the aim of evaluating their influence on the enterotoxic properties of Y. enterocolitica strains.ResultsTwo genotypes (A and G) of the examined nucleotide sequence and some variations were detected in the HRM analysis. A phylogenetic analysis of 10 genotype A nucleotide sequences revealed 100% similarity with the Yersinia enterocolitica subsp. enterocolitica 8081 genome NCBI Acc. No. AM286415. An analysis of 10 genotype G nucleotide sequences and 3 variations sequences revealed two point mutations in the examined region: transition A3387326G and insertion A in position 3387368. However, no mutations were observed in the coding region of any of the examined ymoA gene fragments. Genotype G was identified in nearly all Y. enterocolitica strains isolated from pigs. Only 4 nucleotide sequences were similar to AM286415 and did not feature point mutations. In case of human Y. enterocolitica strains 31 were classified as belonging to genotype A, the remaining 59 belonged to genotype G and were characterized by the presence of point mutations.ConclusionsNo correlations were observed between enterotoxic properties and the presence of mutations in the ymoA gene region of Y. enterocolitica strains isolated from both humans and pigs.

WITHDRAWN: The Impact of ymoA Gene Expression on Enterotoxin YstA Production by Yersinia Enterocolitica Strains With Different Enterotoxic Properties

“…However, the results of our previous study 23 show that two point mutations in the nucleotide sequence of the ymoA gene, which were detected with the use of the High-Resolution Melting (HRM) method, did not in uence on enterotoxic properties of the examined strains. Mikulskis et al 9 in 1994 presented the mechanism modifying the expression of yst to a silent state.…”

A study of single nucleotide polymorphism in the <i>ystB</i> gene of <i>Yersinia enterocolitica</i> strains isolated from various wild animal species

“…According to best knowledge of the authos', this is one of the few studies where the HRM method was deployed to characterize Y. enterocolitica strains. Bancerz-Kisiel et al [16] used HRM to detect possible mutations in the ymoA (Yersinia Phylogenetic analysis of G1 nucleotide sequences ystB gene Y. enterocolitica strains isolated from wild animals revealed 100% similarity with the Y. enterocolitica D88145.1 sequence. Analysis of G2 nucleotide sequences ystB gene revealed one point mutation in the examined region: transition T111A.…”

“…High-resolution melting (HRM) analysis is an emerging method for the detection of single nucleotide polymorphisms (SNPs) and for SNPs-based genotyping [16,17]. In this method, the reduction in fluorescence is carefully monitored when a PCR product stained with a double-strand-specific fluorescent dye is heated through its melting temperature (T m ) [18].…”

The Most Important Virulence Markers of Yersinia enterocolitica and Their Role during Infection

“…Mikulskis et al [ 83 ] also postulated the inherence of a mechanism switching the yst expression to a silent state, and other authors showed that yst gene presence is not always correlated with YstI production [ 85 , 95 ]. In the study of Bancerz-Kisiel et al [ 96 ], no mutations were found in the coding region of the examined ymoA gene fragments. Two point mutations in the non-coding region have been identified in some of the tested Y. enterocolitica strains, regardless of their enterotoxic properties [ 96 ].…”

“…In the study of Bancerz-Kisiel et al [ 96 ], no mutations were found in the coding region of the examined ymoA gene fragments. Two point mutations in the non-coding region have been identified in some of the tested Y. enterocolitica strains, regardless of their enterotoxic properties [ 96 ]. Hence, the suggestion that ymoA gene mutations influence ystA gene silencing was not confirmed.…”