Title: Detection of Brucella melitensis in goats by conventional and molecular assays

Authors: NK Sandhu, NS Sharma, Paviter Kaur and Y Sandhu

Source: Ruminant Science (2019)-8(1):13-18

Cite this reference as: Sandhu NK, Sharma NS, Kaur Paviter and Sandhu Y (2019). Detection of Brucella melitensis in goats by conventional and molecular assays. Ruminant Science 8(1):13-18.

Abstract

Brucellosis in goats is an economically important disease characterised by abortion, retained placenta, weak offspring and infertility. Though Brucella spp. show a host specificity, but some strains can be transmitted among a variety of animals, including humans. In the present study, a total of six isolates of B. melitensis isolated from 30 clinical samples of fetal stomach contents, vaginal swabs and discharges, placenta, uterine discharges and aborted material of goats were characterized using biochemical and molecular assays. Bruce ladder PCR was employed for detection and species identification of various Brucella isolates to study the prevalence patterns and to check whether the organisms were isolated from their natural hosts or not. By PCR using B4/B5 primer pair, an amplicon size of 223 bp was obtained thus confirming the isolates to be that of Brucella spp. By Bruce Ladder Multiplex PCR, amplified product size of 1682bp, 1071bp, 794 bp, 587bp, 450bp and 152bp was obtained in all the isolates thus confirming all the isolates as B. melitensis. DNA extraction directly from clinical samples of foetal stomach content, vaginal mucus, aborted material (spleen, liver), and cotyledons were carried out with the help of commercially available DNA extraction kit. Of the 30 DNA extracted directly from clinical samples, nine samples were positive for Brucella spp. by PCR using B4/B5 primer pair and amplicon size of 223 bp was obtained. Thus, species identification of Brucella spp. using multiplex assays will further help in understanding the epidemiology of brucellosis for effective control programs.

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