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Updated: Jul 18, 2026

Multi-locus Variable-number Tandem-repeat Analysis of the Fish-pathogenic Bacterium Yersinia ruckeri by Multiplex PCR and Capillary Electrophoresis

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Typing and subtyping of Clostridium difficile isolates by using multiple-locus variable-number tandem-repeat

Renate J van den Berg¹, Inge Schaap, Kate E Templeton

¹Department of Medical Microbiology, Center of Infectious Diseases, Leiden University Medical Center, Leiden, The Netherlands.

Journal of Clinical Microbiology

|December 15, 2006

Summary

A new typing method, multiple-locus variable-number tandem-repeat analysis (MLVA), effectively differentiates Clostridium difficile strains. This highly discriminatory genotyping tool aids in tracking outbreaks and identifying emerging variants of C. difficile.

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Multi-Locus Sequencing for Strain Typing of Toxoplasma gondii

Multi-Locus Sequencing for Strain Typing of Toxoplasma gondii

Published on: May 26, 2026

Area of Science:

Microbiology
Genetics
Epidemiology

Background:

Clostridium difficile is a significant cause of healthcare-associated infections.
Accurate and discriminatory typing methods are crucial for understanding C. difficile transmission and outbreaks.
Existing typing methods may have limitations in resolving closely related strains.

Purpose of the Study:

To develop and validate a novel multiple-locus variable-number tandem-repeat analysis (MLVA) method for Clostridium difficile typing.
To assess the discriminatory power of MLVA for differentiating reference strains and clinical isolates.
To evaluate MLVA's utility in subtyping emerging and outbreak-associated C. difficile strains.

Main Methods:

Development of MLVA based on the genomic sequence of Clostridium difficile strain 630.
Utilized automated fragment analysis and multicolored capillary electrophoresis for high-throughput processing.
Applied MLVA to a diverse collection of reference strains, outbreak isolates (PCR ribotype 027), and toxin A-negative, toxin B-positive isolates (PCR ribotype 017).

Main Results:

MLVA successfully discriminated all tested reference strains, including those from 31 serogroups, 25 toxinotypes, and 7 subtypes of PCR ribotype 001.
Analysis of 28 isolates from PCR ribotype 027 outbreaks revealed 13 distinct clusters, highlighting MLVA's ability to subtype hypervirulent strains.
Twenty-nine PCR ribotype 017 isolates from eight countries formed eight country-specific clusters, demonstrating geographical discrimination.

Conclusions:

Multiple-locus variable-number tandem-repeat analysis (MLVA) is a highly discriminatory genotyping method for Clostridium difficile.
MLVA provides a valuable new tool for the epidemiological surveillance and subtyping of emerging C. difficile variants.
The method's ability to resolve closely related strains aids in outbreak investigations and understanding transmission dynamics.