Document Type : Original Article

Authors

• Masoumeh Noura ¹
• Hossein Kamaladini ¹
• Fatemeh Haddadi ¹
• Mohsen Najimi ²

¹ Department of Biology, Faculty of Sciences, University of Zabol, Zabol, Iran

² Department of Pathobiology, Faculty of Veterinary Medicine, University of Zabol, Zabol, Iran

Abstract

Brucellosis or Malta fever (Mediterranean fever) is an important zoonosis caused by different species of Brucella – a small, Gram-negative, aerobic, non-motile, non-encapsulated, and non-spore-forming coccobacillus. Brucellosis can be easily transmitted to humans by Brucella-contaminated blood, meat, or milk. The lack of an effective tool for vaccination or efficient treatment has necessitated rapid bacterial detection methods for preventing this disease. In this study, we optimized the molecular detection of Brucella through polymerase chain reaction (PCR) and multiplex-PCR. To this end, the Omp31 and BLS genes were amplified, resulting in two fragments of 347 bp and 256 bp, respectively. PCR and multiplex-PCR specificity and sensitivity for genomic DNA were 100% and 0.39 ng/μL, respectively. The detection time of Brucella was less than 2 hours, which is obviously shorter than the identification time of the traditional methods like culture, which usually takes more than a day. Given the high specificity and sensitivity of Brucella detection with these genes through multiplex-PCR, we suggest this approach for evaluating the contamination of livestock in veterinary reference laboratories.

Graphical Abstract

Keywords

• Brucella
• Brucellosis
• BLS gene
• Omp31 gene
• Multiplex polymerase chain reaction
• Polymerase chain reaction

Main Subjects

• Basic Science