Rapid detection of causative agents of Malta fever using polymerase chain reaction

ABSTRACT

Malta fever is a zoonosis that it's causative agent is Brucella species. Although the disease is rarely fatal, but in natural epidemics or probable biologic wars, it's rapid and proper indiscrimination will scathe medical supplies drastically. Drawbacks of traditional diagnostic methods are time consuming, less accurate, less sensitive and contain false positive results. So the aim of this study is to design the PCR method for rapid detection of the organism. Genus specific bcsp target gene was selected for primer designing using Primer express v3.0 software. Specificity of the PCR tests was determined using reactions containing various bacterial negative control genomes. For evaluation of sensitivity and limit of detection, the PCR products were cloned in pTZ57R/T plasmid and used from their serial dilutions. Electrophoresis of bcsp gene PCR product showed a single 150 bp band for B.abortus and B.melitensis. PCR tests using negative control bacterial genomes were not showed detectable bands after gel agarose electrophoresis. The cloning processes were confirmed by PCR and sequencing. This method can be used for genus detection of the pathogen Brucella. and, it reduces time and cost needed for the organisms detection