16- DETECTION OF BRUCELLA SPECIES IN APPARENTLY HEALTHY COWS AND GOATS RAW MILK BY PCR

Abstract

Brucellosis an intracellular pathogen capable of infecting animals and humans.The aim of this study was to identify Brucella spp in apparently healthy cow and goatraw milk samples by a polymerase chain reaction (PCR)-based method. A total of 75cow milk samples(55 direct and 20 indirect) and 50 goat milk samples wereexamined by PCR. To establish a PCR protocol for diagnosis of brucellosis, DNAwas extracted from the milk samples by using a commercial kit. PCR amplificationwas done for detection of Brucella DNA using BCSP31 target gene and IS711 locus.The PCR assay showed that an amplicon of 223 bp was obtained in 28% (21/75) and8% (4/50) samples of cow and goat tested milk using primers (B4/B5) derived fromaBCSP31gene encoding the 31-kDa Brucella abortus antigen. In another PCR, anamplicon of 498 bp was obtained in 100% (21/21) of the brucella genus BCSP31-PCRpositive cow milk samples using Brucella abortus-specific primers derived from alocus adjacent to the 3’-end of IS711, and also an amplicon of 731 bp was produced in100% (8/8) of the BCSP31-PCR positive goat milk samples using Brucellamelitensis-specific primers.PCR positive results for brucella genus and brucellaabortus species were observed in all (100%) cow,s milk samples(n=11) with clinicalhistory of abortion.As a single PCR product, no B. abortus was detected in goat milksamples alsoB. Melitensis was not detected in cow milk samples

Keywords

PCR, Milk, Brucellosis.