Molecular characterization of Brucella melitensis detected in blood and Serum samples of sheep and Goats

Abstract

The incidence of brucellosis was investigated in sheep and goats with a history of abortions. Out of 550 blood samples collected from small ruminants, a total of 113 samples including 44 blood and 69 serum samples tested positive for brucellosis by using i-enzyme linked immunosorbent assay. Further, these samples were subjected to molecular detection of Brucella spp by BCSP31 and IS711/BM polymerase chain reaction (PCR) assays. The reference strains B. abortus S19, B. abortus 544 and B. melitensis Rev 1 were included in PCR assay as controls. The reference strains B. abortus S19 and B. abortus 544 produced amplification product of 223 bp. Moreover, 223 bp amplicon was observed in 12 (27.27%) blood and 14 (20.29%) serum samples confirming the presence of Brucella spp. In IS711/BM PCR B. melitensis Rev 1, 10 (22.73%) blood and 10 (14.49%) serum samples produced amplicons of 731 bp confirming the presence of B. melitensis. Phylogenetic analysis of four IS711/BM gene sequences of B. melitensis detected the ancestral origin of organism. It was inferred that BCSP31 and IS711/BM PCR assays were found useful in direct detection of Brucella organisms from whole blood and serum samples of sheep and goats.