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Identification of invasive Yersinia species using oligonucleotide probes

P Feng1

  • 1Division of Microbiology, Food and Drug Administration, Washington, DC 20204.

Molecular and Cellular Probes
|August 1, 1992
PubMed

Insights

Specific oligonucleotide probes accurately identified invasive Yersinia species. The INV-3 probe detected the inv gene in Yersinia pseudotuberculosis, while the PF-13 probe identified the ail gene in Yersinia enterocolitica.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Genetics

Background:

  • Yersinia species are significant human pathogens.
  • Invasion genes like inv and ail are crucial for Yersinia pathogenicity.
  • Accurate identification of invasive strains is vital for clinical and epidemiological purposes.

Purpose of the Study:

  • To develop and validate specific oligonucleotide probes for detecting invasion genes in Yersinia species.
  • To assess the correlation between the presence of inv and ail genes and the invasive capabilities of Yersinia isolates.
  • To differentiate between pathogenic Yersinia species and other bacteria.

Main Methods:

  • Colony hybridization using specific oligonucleotide probes (INV-3 and PF-13).
  • Southern blot analysis of genomic DNA digested with restriction enzymes.
  • Testing probe specificity against Yersinia and non-Yersinia isolates.
  • Correlation of probe reactivity with HeLa cell invasion assays.

Main Results:

  • The INV-3 probe specifically hybridized with all invasive Yersinia pseudotuberculosis isolates (48/48) targeting the inv gene.
  • The PF-13 probe specifically identified invasive Yersinia enterocolitica strains (36/52) by targeting the ail gene.
  • Neither probe showed cross-reactivity with non-Yersinia species or non-invasive Yersinia strains.
  • Southern analyses confirmed probe specificity to known gene fragments (inv: 4.5 kb Bam HI; ail: 1.2 kb Cla I-Ava I).
  • Probe reactivity strongly correlated with the ability of isolates to invade HeLa cells.

Conclusions:

  • Oligonucleotide probes INV-3 and PF-13 are highly specific and sensitive tools for identifying invasive Yersinia pseudotuberculosis and Yersinia enterocolitica, respectively.
  • These probes provide a rapid and reliable method for detecting key virulence genes associated with bacterial invasion.
  • The developed probes can aid in the accurate diagnosis and epidemiological surveillance of Yersinia infections.

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