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Updated: Mar 3, 2026

Microscopy-based Assays for High-throughput Screening of Host Factors Involved in Brucella Infection of Hela Cells

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Genotyping Brucella canis isolates using a highly discriminatory multilocus variable-number tandem-repeat analysis

Yi Yang^1,2, Yin Wang¹, Elizabeth Poulsen¹

¹Kansas State Veterinary Diagnostic Laboratory, College of Veterinary Medicine, Kansas State University, Manhattan, KS, 66502, USA.

Scientific Reports

|April 23, 2017

Summary

A new MLVA-13Bc method effectively genotypes Brucella canis strains, outperforming previous techniques. This highly discriminatory assay aids in tracking B. canis outbreaks and understanding canine brucellosis epidemiology.

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Published on: June 17, 2019

Area of Science:

Veterinary Microbiology
Molecular Epidemiology
Canine Infectious Diseases

Background:

Brucella canis causes canine brucellosis, a significant zoonotic concern.
Current methods like PCR and standard MLVA struggle to differentiate B. canis strains effectively.
Accurate strain differentiation is crucial for epidemiological investigations and outbreak control.

Purpose of the Study:

To develop and validate a novel, highly discriminatory genotyping method for Brucella canis.
To assess the discriminatory power of the new MLVA-13Bc assay for B. canis isolates.
To apply the method for molecular epidemiological analysis of B. canis strains in the U.S.

Main Methods:

Development of the MLVA-13Bc assay by combining eight novel and five established VNTR loci.
Genotyping of 229 B. canis isolates collected from canine blood samples between 2010 and 2016 across 22 U.S. states.
Calculation of the Hunter-Gaston Discriminatory Index for individual loci and the combined assay.

Main Results:

The MLVA-13Bc method successfully differentiated all 229 B. canis isolates.
Individual VNTR loci showed high discriminatory power (0.516-0.934), with a combined index of 1.000.
Analysis revealed three major clusters (A, B, C) and 10 distinct genotype groups among the isolates.

Conclusions:

The MLVA-13Bc assay is a robust and highly discriminatory tool for Brucella canis genotyping.
This method significantly enhances molecular epidemiological capabilities for B. canis.
It is particularly valuable for tracing contamination sources during B. canis outbreaks.