Molecular detection of some virulence factors genes and biofilm formation in clinical isolates of Yersinia enterocolitica

Document Type : Research Paper

Authors

Department of Biology, College of Education, University of Al-Qadisiyah, Iraq.

10.22103/jab.2025.25502.1726

Abstract

Objective
Yersinia enterocolitica is a foodborne pathogen responsible for gastrointestinal infections, notably diarrhea, with its virulence attributed to genetic factors enabling host cell invasion, immune evasion, and environmental persistence. This investigation aimed to detect the 16S rRNA gene, evaluate virulence-related genes (yadA and hreP), and evaluate biofilm formation in Y. enterocolitica isolates from diarrheal samples gathered in Babylon Province, Iraq, to elucidate their genetic diversity and pathogenic potential.

Materials and Methods
A total of 200 stool samples from patients of varying ages with diarrhea were gathered from multiple hospitals across Babylon Province. Isolates were identified applying standard biochemical experiments and polymerase chain reaction (PCR) targeting the 16S rRNA gene of Y. enterocolitica, followed by gene sequencing. Molecular diagnosis of virulence genes yadA and hreP was carried out applying specific primers. Biofilm formation was evaluated through quantitative assays to identify the isolates’ capability to adhere to surfaces, reflecting their potential for persistence in clinical environments. Sequence data were analyzed applying multiple sequence alignment and phylogenetic tree construction to evaluate genetic relatedness.

Results
Genetic diversity was evaluated via 16S rRNA gene sequencing, revealing high conservation with six single nucleotide polymorphisms (SNPs), primarily transitions, located in non-coding or structurally neutral regions. Iraqi isolates demonstrated close relatedness among some strains but phylogenetic diversity in others, clustering into three major clades: First Genetically Unified (FGU), First New Diverging (FND), and Final Unique (FU). Sequence alignment included two Iraqi sequences (GenBank: PV628219, PV628221) and 17 reference sequences from GenBank, with conserved nucleotides color-coded (A: green, T: red, G: purple, C: blue) and SNPs highlighted. Iraqi strains were highly similar to the reference sequence PV628226, with SNPs at positions 19 and 141 denoting close relatedness. In total, six SNPs were identified, with strains PV628219 and PV628221 each exhibiting four SNPs. Phylogenetic analysis affirmed diverse genetic profiles among Iraqi Y. enterocolitica isolates, with some strains closely related and others more divergent. Virulence genes yadA and hreP were detected in 87.5% of isolates (7/8), suggesting meaningful pathogenic potential. Biofilm formation assays revealed that most isolates exhibited moderate to strong biofilm production, denoting their capacity to persist in clinical settings.

Conclusions
This investigation highlights the genetic diversity and pathogenic potential of Y. enterocolitica isolates from Babylon Province, Iraq. The identification of SNPs in the 16S rRNA gene and the presence of virulence genes yadA and hreP in most isolates underscore their molecular basis for pathogenicity.

Keywords

References

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Agricultural Biotechnology Journal
Volume 17, Issue 3
August 2025
Pages 218-201

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