Development of a diagnostic multiplex polymerase chain reaction microarray assay to detect and differentiate Brucella...

Area of Science:

• Veterinary microbiology
• Molecular diagnostics
• Food safety science

Background:

• Brucellosis is a significant global zoonotic disease causing economic losses and human health issues.
• Accurate and rapid laboratory diagnostics are crucial for controlling Brucellosis in humans, animals, and food production.
• Current diagnostic methods may require improvement for speed and high-throughput analysis.

Purpose of the Study:

• To develop a rapid, multiplex polymerase chain reaction microarray assay for identifying the Brucella genus and differentiating its species.
• To create a high-throughput molecular test for Brucella detection and analysis.

Main Methods:

• A multiplex polymerase chain reaction microarray assay was designed using 11 signature sequences and redundant oligonucleotide probes.
• Genus-specific gene targets (bcsp31, perA, cgs, omp2b) and species-specific chromosomal regions were utilized.
• The assay was validated using Brucella reference strains and 102 field isolates.

Main Results:

• The Brucella ArrayTube assay accurately identified Brucella reference strains and field isolates based on hybridization patterns.
• Genus identification was unambiguous for all tested isolates.
• Species differentiation was limited for specific groups, including B. suis bv 3/4/B. canis and B. neotomae/B. microti.

Conclusions:

• The Brucella ArrayTube assay offers an easy-to-handle, molecular method for high-throughput, parallel analysis of Brucella.
• This assay facilitates rapid identification of the Brucella genus.
• Further refinement may be needed for precise species differentiation within certain Brucella groups.