Typing and clustering of Yersinia pseudotuberculosis isolates by restriction fragment length polymorphism analysis
E Voskresenskaya1, C Savin2, A Leclercq2
1Yersinia Research Unit, National Reference Laboratory, WHO Collaborating Center for Yersinia, Institut Pasteur, Paris, France Laboratory of Bacterial Respiratory Infections, Institut Pasteur, Saint Petersburg, Russia.
Restriction fragment length polymorphism of insertion sequence hybridization patterns (IS-RFLP) is an effective method for typing Yersinia pseudotuberculosis. This molecular typing technique, particularly using two insertion sequences (2IS-RFLP), offers high discriminatory power for global isolates.
Area of Science:
- Microbiology
- Molecular Biology
- Infectious Diseases
Background:
- Yersinia pseudotuberculosis is an enteropathogen with an animal reservoir, causing human infections primarily in temperate and cold climates.
- Previous Y. pseudotuberculosis subtyping methods were limited by small sample sizes and geographical restrictions.
- There is a need for efficient and globally applicable molecular typing methods for Y. pseudotuberculosis.
Purpose of the Study:
- To evaluate the typing efficiency of insertion sequence-based restriction fragment length polymorphism (IS-RFLP) compared to serotyping, ribotyping, and multilocus sequence typing (MLST).
- To assess the discriminatory power and clustering capacity of IS-RFLP for global Y. pseudotuberculosis isolates.
- To determine the utility of IS-RFLP for analyzing the population structure of Y. pseudotuberculosis.
Main Methods:
- Typing of 80 global Y. pseudotuberculosis strains using IS-RFLP with IS285 and IS1541, alongside serotyping, ribotyping, and MLST.
- Calculation of discriminatory index (DI) for each typing method.
- Analysis of the clustering power of IS-RFLP to group strains with similar markers.
Main Results:
- IS285-RFLP (DI=0.939) and IS1541-RFLP (DI=0.980) demonstrated higher discriminatory power than serotyping (DI=0.857), MLST (DI=0.861), and ribotyping (DI=0.944).
- The combined 2IS-RFLP method achieved a superior DI of 0.998.
- 2IS-RFLP effectively grouped strains with shared genotypic and phenotypic characteristics, revealing robust population structures.
Conclusions:
- IS-RFLP, particularly using IS1541 or the combination of two insertion sequences (2IS-RFLP), is a powerful and efficient tool for molecular typing of global Y. pseudotuberculosis isolates.
- IS-RFLP provides well-resolved banding patterns suitable for reliable strain comparison.
- The method demonstrates significant potential for analyzing the population structure of Y. pseudotuberculosis species.
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