Development of A Per Gene-Based PCR Assay for the Detection of Brucella Species in Spiked Milk Samples

Brucellosis is a significant zoonotic disease with major economic implications, particularly in livestock. Traditional diagnostic methods, such as bacterial culture and serology, are often time-consuming and less sensitive. In this study, a PCR assay targeting the per gene was developed. The assay demonstrated 100% sensitivity in detecting Brucella DNAacross various isolates and showed no cross-reactivity with non-Brucella species. It was capable of detecting as little as 38.3 pg of Brucella genomic DNA and 4.8 × 10² CFU/mL in spiked milk samples, suggesting its high sensitivity and specificity. The developed assay offered a rapid, reliable method for Brucella detection, particularly useful for outbreak investigations and zoonotic risk assessment. Future research should focus on optimizing sample preparation and expanding testing across diverse sample types to further enhance the assay’s applicability in field conditions.