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Multiplex Multilocus Variable-Number Tandem-Repeat Analysis for Typing of Pandemic Vibrio parahaemolyticus O1:KUT

Sutima Preeprem1, Kamonnut Singkhamanan2, Mitsuaki Nishibuchi3

  • 11 Department of Microbiology, Faculty of Science, Prince of Songkla University , Songkhla, Thailand .

Foodborne Pathogens and Disease
|October 17, 2018
PubMed

Insights

A new multiplex multilocus variable-number tandem-repeat analysis (MLVA) assay effectively types pandemic Vibrio parahaemolyticus strains, including difficult-to-trace O1:KUT isolates. This molecular method offers superior discrimination compared to traditional techniques for tracking foodborne illness outbreaks.

Area of Science:

  • Microbiology
  • Molecular Epidemiology
  • Food Safety

Background:

  • Pandemic Vibrio parahaemolyticus O3:K6 emerged in 1996, causing global gastroenteritis with serovariants like O1:KUT.
  • Limitations in K typing antisera hinder epidemiological tracking of O1:KUT isolates.
  • Effective molecular typing methods are crucial for discriminating pandemic V. parahaemolyticus strains.

Purpose of the Study:

  • To develop a multiplex multilocus variable-number tandem-repeat analysis (MLVA) assay.
  • To enhance the typing of pandemic V. parahaemolyticus, specifically including O1:KUT isolates.
  • To improve molecular discrimination of V. parahaemolyticus strains for epidemiological investigations.

Main Methods:

  • Development of a multiplex MLVA assay targeting four variable number tandem repeat loci.
  • Analysis of 46 pandemic V. parahaemolyticus isolates, including O1:KUT, O1:K25, and O3:K6 serotypes.
  • Comparison of MLVA results with pulsed-field gel electrophoresis (PFGE) typing.

Main Results:

  • The multiplex MLVA assay generated 38 distinct profiles, significantly higher than the 16 types from PFGE.
  • MLVA successfully differentiated 12 O1:KUT isolates with identical PFGE patterns into unique profiles.
  • The developed MLVA assay demonstrated a high discriminatory power (D=0.99).

Conclusions:

  • Multiplex MLVA is a powerful tool for typing pandemic V. parahaemolyticus.
  • The assay provides superior strain discrimination compared to PFGE, particularly for O1:KUT isolates.
  • This method enhances epidemiological investigations by enabling precise tracking of V. parahaemolyticus sources.
Keywords:
MLVAPFGEVNTRmolecular epidemiology

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