Development of a multiplex polymerase chain reaction assay for differentiation of field strain isolates and vaccine strains S19 and RB51 of Brucella in Iran

Document Type : Full paper (Original article)


1 Department of Clinical Sciences, School of Veterinary Medicine, Shiraz University, Shiraz, Iran

2 Department of Clinical Sciences, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran

3 Department of Microbiology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran

4 Department of Brucella Vaccine Research and Production, Razi Vaccine and Serum Research Institute, Karadj, Iran


Bovine brucellosis is a zoonotic disease distributed worldwide and characterized by abortion and reduced
fertility in cows. Since brucellosis eradication programme in Iran uses vaccination, test, slaughter and
quarantine as control measures, it is essential to distinguish vaccine strains from strains that cause infections
among vaccinated cattle herds. We developed and evaluated a multiplex polymerase chain reaction (PCR)
assay to identify and differentiate the vaccine strains from wild field isolates, including all Brucella types
usually found in cattle in Iran. Two pair primers were used to amplify eri and wbo regions of DNA sequences
those strain-specific targets for B. abortus S19 and RB51 vaccine strains. This multiplex PCR method
evaluated with DNA from reference strains, two vaccine strains and 29 field strains of Brucella. The results
showed that the multiplex PCR can differentiate Brucella isolates into three categories: strain 19 (S19), strain
RB51 and field strains. This PCR assay was successfully used, compared with traditional method to
differentiate of S19 and RB51 from field Brucella isolates