Evaluation of two PCR techniques for detection of Brucella DNA in Contaminated Serum Sample

Year 2015, Volume: 36 Issue: 3, 1706 - 1716, 13.05.2015

K. Moghadas , E. Ghezelsofla B. Zamani M. Moghni H. Aghamolaee F. Moosavi H. Karimzade

Abstract

Abstract. Brucellosis is a major health problem worldwide.Currently the diagnosis of this zoonosis is based on microbiological and serological tests.PCR has been used to detect DNA from Brucella. PCR techniques and extraction procedures have been previously published for Brucella detection.But only a few of these primers have been used in human samples.and only a few study has been carried out to compare sensitivity between them.In this study,two sets of primers amplifying two different regions of the Brucella genome were compared for detection of Brucella DNA in Contaminated Serum Sample PCR assay to conclude which is most suitable for the clinical diagnostic laboratory.These two pairs of primers amplify:i)a sequence 16SrRNA of B.abortus(F4/R2),and(ii)a gene encoding an outer membrane protein(omp-2)(JPF/JPR).The two primers assayed showed a difference in sensitivity for detecting Brucella DNA,ranging between 5Pg and 50Pg for contaminated serum samples.Therefore,the sensitivity of PCR using F4/R2 primers was greater than the PCR using JPF/JPR primers.

Keywords

brucellosis, PCR, contaminated serum, omp-2, 16S rRNA

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