Detection of Brucella Species from Seropositive Animal Blood Clots using real time PCR assay - Abstract
Brucellosis is a zoonotic disease transmitted from animal to human. It is economically important diseasecausing abortion, sterility and still birth in adult animals. The diseases can be diagnosed using serological, bacteriological and Molecular methods.The study was conducted to detect Brucella species from blood clot of C-ELISA positive samples using Real Time PCR. Blood sample collected and blood clot separated from collected blood samples. Serum sampleswere screened using Rose Bengal Plate Test finally confirmed by C-ELISA. Blood clot of C-ELISA positive blood clot of cattle and small ruminant subjected to PCR assay to detect Brucella species. Bacterial DNA was extracted according to Qiagen DNA extraction kit standard procedure and amplified. Species specific primers for B. abortus and B. meltensis were used to detect Brucella species from extracted DNA. Brucella abortus was detected from blood clot of cattle but from blood clot of small ruminant neither B. meltensisnor B. abortus detected. Molecular methods accompanied with screening and confirmatory serological testsare more suitable method to detect Brucella species than bacteriological method in developing countries; where Biosafety equipment or materials are not adequately available for safety of lab personnel’s. Therefore, laboratories should be capacitated for molecular method of diseases diagnosis and investigation.